2

IAOe?IIAEUeIA AEAAeAI?ss IAOE OE?A?IE

?inoeooo iieaeoey?ii? a?ieia?? ? aaiaoeee

OAeE 57.085.23, 577.122.5, 577.217.535

Iaa?ooeueeee Ai?en Na?a?eiae/

Ooieoe?iiaeueia eiiia?oiaioae?caoe?y aia?aoo a?eeiaiai neioaco o aeueo
aoea??io

03.00.03 — iieaeoey?ia a?ieia?y

Aaoi?aoa?ao

aeena?oaoe?? ia caeiaoooy iaoeiaiai nooiaiy

aeieoi?a a?ieia?/ieo iaoe

Ee?a — 1999

Aeena?oaoe??th ? ?oeiien.

?iaioo aeeiiaii o a?aeae?e? iaoaieci?a o?aineyoe?? aaiaoe/ii?
?ioi?iaoe?? ?inoeoooo iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie

Iaoeiaee eiinoeueoaio:

aeieoi? a?ieia?/ieo iaoe, i?ioani?, aeaaeai?e IAI Oe?a?ie ?euenueea A.A.

?inoeooo iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie, caa. a?aeae?eii.

Io?oe?ei? iiiiaioe:

aeieo.a?ie.iaoe, i?ioani? Aeiia?aaeiaa ?.I., eaoaae?a a?io?i??
Ee?anueeiai oi?aa?neoaoo ?i. O.A.Oaa/aiea, i?ioani?

aeieo.a?ie.iaoe, no.i.nio?. Aaee?i A.I., caa?aeoaa/ a?aeae?eo
a?i?iaeaia??? ?inoeoooo a?ii?aai?/ii? o?i?? oa iaooio?i?? IAI Oe?a?ie

aeieo.a?ie.iaoe, no.i.nio?. Eiceia A.I., i?ia?aeiee iaoeiaee
ni?a?ia?oiee a?aeae?eo aiceiieia?? a?eeiaiai neioaco ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie

I?ia?aeia onoaiiaa: ?inoeooo a?io?i?? ?i. I.A. Iaeeaae?ia IAI Oe?a?ie,
i.Ee?a

Caoeno a?aeaoaeaoueny “ 20 ” ea?oiy 1999 ?ieo i 10-00 aiaeei?

ia can?aeaii? niaoe?ae?ciaaii? a/aii? ?aaee Ae 26.237.01 ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie ca aae?anith: 252143,
Ee?a-143, aoe. Caaieioiiai, 150

C aeena?oaoe??th iiaeia iciaeiieoeny o a?ae?ioaoe? ?inoeoooo
iieaeoey?ii? a?ieia?? ? aaiaoeee IAI Oe?a?ie.

Aaoi?aoa?ao ?ic?neaiee 13 aa?aciy 1999 ?ieo

A/aiee nae?aoa? niaoe?ae?ciaaii? a/aii? ?aaee,

eaiaeeaeao a?ieia?/ieo iaoe Eoeao E.E.

CAAAEUeIA OA?AEOA?ENOEEA ?IAIOE

Aeooaeuei?noue i?iaeaie. A?ineioac a?eea ?aae?co? aaiaoe/io
?ioi?iaoe?th, caeeaaeaio a AeIE, aeey caaacia/aiiy aeeoo?ae?yeueiino?
ee?oeie. Ianueiaiaei? noaei i/aaeaeiei cia/ia oneeaaeiaiiy i?ioeano
a?eeiaiai neioaco o aeueo aoea??io a ii??aiyii? c i?iea??ioaie. Aeey
i?aeo?eiaiiy aoaeoeaiino? o?aineyoe?? a cia/ii ca?eueoaieo ca ia’?iii
ee?oeiao aeueo aoea??io ? iaiao?aeiei ye?nii iiaee ??aaiue i?aai?caoe??,
ooieoe?iioaaiiy oa ?aaoeyoe?? aeoeaiino? o?aineyoe?eiiai aia?aoo.
Iai/iei na?aeioeoaii ia ei?enoue eiai ?nioaaiiy iiaea aooe ia??aiii??iee
?iciiae?e, aai eiiia?oiaioae?caoe?y, eiiiiiaio?a a?eie-neioacoth/iai
aia?aoo a aoea??ioe/i?e ee?oei? (Ryazanov et al, 1987). Ia /an ii/aoeo
iaoeo aeine?aeaeaiue aoei io?eiaii ?yae iai?yieo aeieac?a
eiiia?oiaioae?caoe?? aia?aoo o?aineyoe??, cie?aia, aeyaeaii
nii??aeiai?noue ai?iiaoeee-o?IE neioaoac aei ?eainii (Irvin et al, 1972,
Ovchinnikov et al, 1978), ooai?aiiy aenieiiieaeoey?ieo eiiieaen?a
ai?iiaoeee-o?IE neioaoac (Deutscher, 1984), ni?aeieae?caoe?y ??cieo
eiiiiiaio?a aia?aoo o?aineyoe?? a aoea??ioe/ieo ee?oeiao (Howe et al,
1984, Shestakova et al, 1993). An? oe? aeai?, oi/a e na?ae/eee ia
ei?enoue no?oeoo?ii?, noaoe/ii? eiiia?oiaioae?caoe?? a?eeiaiai neioaco o
aeueo aoea??io, aea ia aeaaaee i?yei? ?ioi?iaoe?? i?i iiaeeeao
ooieoe?iiaeueio ?ieue oeueiai yaeua.

Iaeiei ?c iniiaieo i?iya?a ooieoe?iiaeueiiai cia/aiiy
eiiia?oiaioae?caoe?? iiaea aooe /aiae?ia, aai i?yia ia?aaea/a i?ii?aeieo
i?iaeoeo?a aeaiiai iaoaaie?/iiai oeyoo a?ae iaeiiai eiiiiiaioa nenoaie
aei ?ioiai aac aeenioe?aoe?? a ?ic/ei (Srere, 1987). A?aeiii, ui oaeee
iaoai?ci ia? i?noea aeey aeayeeo iaoaaie?/ieo i?ioean?a, i?ioa
aenia?eiaioaeueieo aeaieo uiaei eiai ?ie? a a?ineioac? a?eea ia /an
ii/aoeo iaoeo aeine?aeaeaiue io?eiaii ia aoei (Nie?ei, 1989). E??i oiai,
aaaeeeaei na?aeioeoaii iioaioe?eiiai ?aaoeyoi?iiai cia/aiiy
eiiia?oiaioae?caoe?? iiaea aooe ci?ia aioo??oiueiee?oeiii? eieae?caoe??
eiiiiiaio?a o?aineyoe?eiiai aia?aoo a caeaaeiino? a?ae ciai?oi?o oiia.

Ioaea, ine?eueee aac c’ynoaaiiy ooieoe?iiaeueii? ?ie?
eiiia?oiaioae?caoe?? iaiiaeeeaa iiaoaeoaaiiy caaaeueii? iiaeae?
i?aai?caoe??, ooieoe?iioaaiiy oa ?aaoeyoe?? a?ineioaco a?eea o aeueo
aoea??io, oi aeaoaeueia aenia?eiaioaeueia ? oai?aoe/ia ?ic?iaea
eiioeaioe?? ooieoe?iiaeueii? eiiia?oiaioae?caoe?? aia?aoo o?aineyoe?? ?
iaei??th c iaea?eueo aeooaeueieo ooiaeaiaioaeueieo i?iaeai no/anii?
iieaeoey?ii? a?ieia?? a?eeiaiai neioaco.

Ca’ycie ?iaioe c iaoeiaeie i?ia?aiaie, ieaiaie, oaiaie. Aeena?oaoe?y
a?aeiia?aea? iniiaiiio ieaio iaoeiai-aeine?aeieo ?ia?o a?aeae?eo
iaoai?ci?a o?aineyoe?? aaiaoe/ii? ?ioi?iaoe?? ?inoeoooo iieaeoey?ii?
a?ieia?? ? aaiaoeee IAI Oe?a?ie (athaeaeaoi? oaie ? 2.2.4.9 “Aea/aiiy
noaae?? aeiiaaoe?? a?ineioaco a?eea o aoea??io” oa “Iieaeoey?i? iniiae
eiiia?oiaioae?caoe?? a?eeiaiai neioaco aeueo aoea??io”). ?iaioa
aeeiioaaeanue oaeiae i?e ni?eyii? io?eiaieo ia eiieo?nieo canaaeao
a?o/eciyieo oa i?aeia?iaeieo i?iaeo?a: ? 5.2/130 “Eiiia?oiaioae?caoe?y
ye iaeei c iaoaieci?a ?aaoeyoe?? a?eeiaiai neioaco” AeEIO Oe?a?ie, ?
5.4/61 “Iaeaiii?/i? aca?iiae?? eiiiiiaio?a a?eie-neioacoth/iai aia?aoo
aeueo aoea??io” ? ? 5.4/73 “Eieae?caoe?y oaeoi??a aeiiaaoe?? o?aineyoe??
o o?a?iaeanoao ethaeeie. Aieea noaio ooieoe?iiaeueii? aeoeaiino?
o?aineyoe?eiiai aia?aoo oa oac ee?oeiiiai oeeeeo” OOAe I?i?noa?noaa
iaoee oa oaoiieia?e Oe?a?ie, a?aioa INTAS ? 96-1594 “the organization
and regulation of EF-1-mediated tRNA/aminoacyl-tRNA channeling in
eukaryotic protein synthesis”, ei?ioei/anieo a?aio?a FEBS oa EMBO.

Iaoa ? caaea/? aeine?aeaeaiiy. Iaoith aeine?aeaeaiiy aoei ?ic?iaeoe oa
aenia?eiaioaeueii iaa?oiooaaoe eiioeaioe?th ooieoe?iiaeueii?
eiiia?oiaioae?caoe?? aia?aoo a?eeiaiai neioaco, aecia/eoe aaaeeea?noue
oaiiiaio eiiia?oiaioae?caoe?? aeey i?aai?caoe??, ooieoe?iioaaiiy oa
?aaoeyoe?? i?ioeano o?aineyoe?? a ee?oeiao aeueo aoea??io.

Aeoiaey/e c iaoe ?iaioe, aoei noi?ioeueiaaii oae? iniiai? caaea/?:

n ?ic?iaeoe iaoiaee ia?oi?aoe?? oeeoiieaciaoe/ii? iaia?aie ee?oei aeueo
aoea??io, ye? aeicaieythoue caa?aaoe aenieo aeoeai?noue
ia?iaaa?e?ciaaieo ee?oei a o?aineyoe??;

aeine?aeeoe eiiia?oiaioae?caoe?th oa /aiae?ia ai?iiaoeee-o?IE
aaciina?aaeiuei a ee?oeiao nnaaoe?a;

aecia/eoe aieea oinoie?aaoeio ia eieae?caoe?th ??cieo noaiaeeieoeue
oaeoi?a aeiiaaoe?? 1 (EF-1I) o o?a?iaeanoao ethaeeie;

?ic?iaeoe iaoiaee io?eiaiiy aenieii/euaieo i?aia?ao?a oaeoi?a
aeiiaaoe?? 1 (EF-1), oai?eaeai?e-o?IE neioaoace oa eiiieaeno aae?e-o?IE
neioaoace c EF-1H ? aea/eoe aca?iiae?th EF-1 c ??cieie ai?iiaoeee-o?IE
neioaoacaie;

aeine?aeeoe iaeaiii?/iee eiiieaen EF-1*GDP*o?IE, cie?aia aecia/eoe
ae?eyiee o?IE, ye? i?eeiathoue o/anoue o aca?iiae?? ?c oaeoi?ii, oa
iiaeeea?noue eiioi?iaoe?eieo ci?i eiiiiiaio?a eiiieaeno.

Iaoeiaa iiaecia iaea?aeaieo ?acoeueoao?a. Aia?oa aaciina?aaeiuei a
ee?oeiao nnaaoe?a iieacaii eiiia?oiaioae?caoe?th oa /aiae?ia
ai?iiaoeee-o?IE i?ae /an o?aineyoe?? i?IE.

Aenia?eiaioaeueii aeiaaaeaii ?nioaaiiy ia?aaeaa/oaaii? a?iioacith
/aiae?iaa o?IE/ai?iiaoeee-o?IE ooieoe?iiaeueii? aca?iiae??
ai?iiaoeee-o?IE neioaoac ?c EF-1 oa iieacaii ??ci? oeyoe ?? cae?eniaiiy.

Aia?oa i?iaeaiiino?iaaii ooai?aiiy iaeaiii?/iiai eiiieaeno EF-1*GDP*o?IE
? cai?iiiiiaaia ooieoe?y oaeiai eiiieaeno, ui iieyaa? a o?ainii?ooaaii?
o?IE a?ae ?eainiie aei ai?iiaoeee-o?IE neioaoace, a oaeiae iieacaii
oi?ioaaiiy /aoaa?oeiiiai eiiieaeno EF-1*GDP*o?IE*ai?iiaoeee-o?IE
neioaoaca.

Iieacaia iiaeeea?noue ci?ie aioo??oiueiee?oeiii? eieae?caoe??
eiiiiiaio?a aia?aoo o?aineyoe?? a caeaaeiino? a?ae ciai?oi?o oiia, ui
na?ae/eoue i?i eiia??i?noue ?nioaaiiy iiaiai ??aiy ?aaoeyoe?? a?eeiaiai
neioaco ca ?aooiie eiai eiiia?oiaioae?caoe?? — aeaeiiia?oiaioae?caoe??.
Aeyaeaii ?ia?a?oi?iee aieea iaaeoeaieo eiioi?ia??a o?IE, ye?
ooai?ththoueny a i?aai?ci? ca iani?eyoeeaeo oiia, ia o?aineyoe?th i?IE,
ui ? iiaei na?aeioeoaii ia ei?enoue ?o o/ano? a ?aaoeyoe?? a?eeiaiai
neioaco a ee?oeiao nnaaoe?a.

Aia?oa ?ic?iaeaii noaio /aiae?iaa o?IE/ai?iiaoeee-o?IE, yea aeaoaeueii
iieno? ??ci? aa??aioe ia?aaea/? o?IE i?ae eiiiiiaioaie aia?aoo
o?aineyoe?? i?ae /an a?eeiaiai neioaco. Cie?aia, aenoiooi i?eiouaiiy i?i
i?ioeeaaeii iai?aaeaio ae?th eiiieaen?a EF-1*GDP e EF-1*GTP a
ia?aianaii? o?IE a?ae ?eainiie aei ai?iiaoeee-o?IE neioaoace oa
ai?iiaoeee-o?IE a?ae neioaoace aei ?eainiie a?aeiia?aeii.

A?oiooth/enue ia io?eiaieo aeaieo, aia?oa i?aaenoaaeaii eiioeaioe?th
ooieoe?iiaeueii? eiiia?oiaioae?caoe?? aia?aoo a?eeiaiai neioaco o aeueo
aoea??io oa aenoiooi a?iioaoe/io iiaeaeue no?oeoo?ii-ooieoe?iiaeueii?
i?aai?caoe?? o?aineyoe?eiiai eiiia?oiaioo.

Oai?aoe/ia oa i?aeoe/ia oe?ii?noue iaea?aeaieo ?acoeueoao?a. Iniiaiei
?acoeueoaoii aeeiiaiiy aeaii? ?iaioe noaea ?ic?iaea eiioeaioe??
ooieoe?iiaeueii? eiiia?oiaioae?caoe?? o?aineyoe?? o aeueo aoea??io, ui ?
?noioiei aeeaaeii o ?icaeoie oai??? aoea??ioe/iiai a?eeiaiai neioaco ?
ia? caaaeueiia?ieia?/ia cia/aiiy o ?icoi?ii? ooiaeaiaioaeueieo iniia
aeeoo?ae?yeueiino? ee?oeie. ?acoeueoaoe aeine?aeaeaiue oe?iei
oeeoothoueny i?aeia?iaeiith iaoeiaith ni?eueiioith oa noai?ththoue
i?aea?oioy aeey iiaeaeueoeo aeine?aeaeaiue a?ineioaco a?eea oa /aiae?iaa
iaoaaie?o?a. Io?eiai? ?acoeueoaoe ?aeiiaiaeothoueny aeey aeei?enoaiiy a
caaaeueiiio eo?n? “Iieaeoey?ia a?ieia?y” oa niaoeeo?n? “A?ineioac a?eea”
aeey nooaeaio?a oi?aa?neoao?a c? niaoe?aeueiino? “a?io?i?y” oa
“iieaeoey?ia a?ieia?y”. Io?eiai? ca ?ic?iaeaiei iaie iaoiaeii i?aia?aoe
oaeoi?a aeiiaaoe?? 1 aeei?enoiaothoueny aeey aea/aiiy ??cieo aniaeo?a
a?eeiaiai neioaco a a?oi? o?ceee ioeeaii?ioa?i?a ?inoeoooo A?eea,
Iou?ii, ?in?y, o a?aeae?e? iaaee/ii? a?io?i?? Oi?aa?neoaoo i.Eaeaeai,
I?aea?eaiaee, a Eaai?aoi??? iieaeoey?ii? aiceiieia?? CNRS,
AE?o-nth?-?aao, O?aioe?y, oa a Oeaio?? Caei?ia’y Oi?aa?neoaoo
Eiiaeo?eooa, Oa?i?iaoii, NOA.

Iniaenoee aianie caeiaoaa/a. Iia? oai?aoe/i? oa aenia?eiaioaeuei? ?aea?,
ui ?ic?iaeyeeny a aeena?oaoe??, iaeaaeaoue aaoi?o. Aenia?eiaioe, ui
iienai? a ?icae?eao 3, 4 oa 6, cae?eniaii iniaenoi aaoi?ii. Iaeei
aenia?eiaio a ?iaio? ?c ia?iaaa?e?ciaaieie ee?oeiaie aeeiiaii a o?niiio
ni?a?ia?oieoeoa? c ?.Noaioeueii?nii. O aeeiiaii? aenia?eiaio?a, ui
i?aaenoaaeai? a ?icae?e? 5, e??i aaoi?a, i?eeiaee o/anoue ni?a?ia?oieee
a?aeae?eo iaoaieci?a o?aineyoe?? aaiaoe/ii? ?ioi?iaoe?? ?IA?A IAIO
O.A.Aoaeeaae/, C.I.Iao?ooaiei ? A.O.Oaeae. Aeine?aeaeaiiy aiaeiaaiii?
oeoi?anoeaioe?? EF-1 aeeiiaii a o?niiio ni?a?ia?oieoeoa? c I.N.
Eaaeio?iei. Io?eiai? a ni?a?ia?oieoeoa? ?acoeueoaoe iioae?eiaaii ye
ni?euei? iaoeia? noaoo?.

Ai?iaaoe?y ?acoeueoao?a aeena?oaoe??. ?acoeueoaoe aeena?oaoe?? aoee
ii?eethaeiai? ia Oeie?-eiioa?aioe?? “No?oeoo?a e ooieoeey aeiiieeia?ia”,
Euea?a, 1988; VI Neiiic?oi? ”Macromolecules in functioning cell”
USSR-Italy, Eai?ia?aae, 1988; I?aeia?iaeiiio neiiic?oi? “Regulation of
translation”, ?eaa, 1989; 14 ?iai/?e Ia?aae? c o?IE, Iiciaiue, Iieueua,
1991; Aeine?aeieoeuee?e Ai?aeiiianuee?e eiioa?aioe?? “Iae?iiieaeoey?ia
i?aai?caoe?y oa ooieoe?y ee?oei”, Ienia?ae, NOA, 1995; 16 ?iai/?e
Ia?aae? c o?IE, Iaae?nii, NOA, 1995; I?aeia?iaei?e eiioa?aioe??
“Frontiers in Translation”, A?eoi??y, Eaiaaea, 1995; 17 ?iai/?e Ia?aae?
c o?IE, O?aa, ssiii?y, 1997; 17 I?aeia?iaeiiio Eiia?an? c A?io?i?? oa
Iieaeoey?ii? A?ieia??, Nai-O?aioe?nei, NOA, 1997; I?aeia?iaei?e
eiioa?aioe?? “Iieaeoey?iay aaiaoeea e aeioaoiieiaey”, I?inuee,
A?ea?onue, 1998; ?iai/?e ia?aae? c? no?oeoo?e oa ooieoe??
ai?iiaoeee-o?IE neioaoac, I?oaeuea??, O?aioe?y, 1998.

Iaoeia? ?acoeueoaoe aeiiia?aeaeeny oaeiae ia nai?ia?ao a?aeae?eo
iaoai?ci?a o?aineyoe?? aaiaoe/ii? ?ioi?iaoe?? ?IA?A IAI Oe?a?ie,
a?aeae?eo a?io?i?? Oi?aa?neoaoo ooaoo Eiiaeo?eoo, NOA, eaai?aoi???
iaaee/ii? aaiaoeee Eaeaeainueeiai Oi?aa?neoaoo, I?aea?eaiaee, ?inoeoooo
iieaeoey?ii? a?ieia?? Oi?aa?neoaoo A?ooai-Oa?aeaea, I?ia//eia,
Eaai?aoi??? iieaeoey?ii? aiceiieia?? CNRS, O?aioe?y, a?oie o?ceee
ioeeaii?ioa?i?a ?inoeoooo A?eea, ?in?y.

Ioae?eaoe??. Iaoa??aee aeena?oaoe?eii? ?iaioe a iiaiiio ia’?i?
iaae?oeiaai? a 27 ioae?eaoe?yo, c ieo 18 noaoae o a?o/eciyieo oa
i?aeia?iaeieo oaoiaeo iaoeiaeo aeaeaiiyo.

No?oeoo?a oa ianya ?iaioe. Aeena?oaoe?y neeaaea?oueny c? anooio, iaeyaeo
e?oa?aoo?e, iieno iaoa??ae?a oa iaoiae?a aeine?aeaeaiue, ?acoeueoao?a oa
?o iaaiai?aiiy, caaaeueieo i?aenoie?a, aeniiae?a oa nieneo e?oa?aoo?e,
yeee aeeth/a? 371 iineeaiiy. ?iaioa aeeeaaeaia ia 336 noi??ieao
iaoeiiieniiai oaenoo ? i?noeoue 74 ?enoiee oa 4 oaaeeoe?.

INIIAIEE CI?NO

Iaoa??aee oa iaoiaee. Ee?oeie y?/ieea eeoaenueeiai oii’y/ea (NII)
iaaeaii Ae.Iaeoa?ii (NOA). Ia?aeiio eoeueoo?o o?a?iaeano?a ethaeeie VH
25 (15-18 ianaae?) aoea io?eiaia a?ae ss.Aeaeea (I?aea?eaiaee). o?IE c
ia/?iee e?iey io?eioaaee i?yiith aeai?ioa?i?caoe??th aiiiaaiaoa oeaieie
oaiieii oa o?iiaoia?ao??th ia DEAE-oeaetheic?, ye iienaii iaie ?ai?oa
(Iaa?ooeeee e ae?., 1985). 40S ? 60S ?eainiii? noa/anoeiee io?eioaaee,
ye iienaii iaie ?ai?oa (Iaa?ooeeee, 1981). Ai?iiaoeee-o?IE a
i?aia?aoeai?e e?eueeino? aoea neioaciaaia ye iienaii (Yang et al, 1971).
o?IEPhe e o?IELeu aoee i/euai? c noia?ii? o?IE ia/?iee e?iey
o?iiaoia?ao??th a nenoai? HPLC Gold ia eieiioe? Hypersil 5C4 (Beckman).
32P-i?/aiiy o?IE i?iaiaeeee ca?aeii (Silberclang et al, 1977). I/euaiiy
EF-1 cae?enithaaee ca ?ic?iaeaiei iaie iaoiaeii, yeee neeaaea?oueny c
oaeeo aoai?a: aaeueo?eueo?aoe?y ia naoae?ee? S-400, o?iiaoia?ao?y ia
DEAE-oeaetheic? oa SP-naoa?ic?. *enoioa i?aia?aoo EF-1 noaiiaeea a?eueo
i?ae 90%. I?aia?ao a?eea iaa aenieo aeoeai?noue a iai?i? GDP/3H-GDP,
eaoae?c? ca’ycoaaiiy oai?eaeai?e-o?IE c iie?(U)-cai?ia?aiiaaieie
?eainiiaie oa neioac? iie?(?he) ia iie?(U)-cai?ia?aiiaaieo ?eainiiao.
I/euaiiy oai?eaeai?e-o?IE neioaoace cae?enithaaee ca?aeii ?c
cai?iiiiiaaiith iaie iaoiaeeeith o?aeoe?iioaaiiyi
iinoi?oioiiae??aeueiiai noia?iaoaioo IAA6000, o?iiaoia?ao??th ia
a?ae?ien?aiaoeo? oa oinoioeaeetheic?, aaeueo?eueo?aoe??th ia Sephacryl
S-300. *enoioa i?aia?aoo oai?eaeai?e-o?IE neioaoace noaiiaeea a?eueo
i?ae 80%, ca?aeii aeaeo?ioi?aco a aeaiaoo?oth/eo oiiaao. I/euaiiy
eiiieaeno aae?e-o?IE neioaoace ? EF-1H c ia/?iee e?ie?a i?iaiaeeee ca
oaeith noaiith: o?aeoe?iioaaiiy IAA6000, aaeueo?eueo?aoe?y ia BioGel
A5m, o?iiaoia?ao?y ia o?IE-naoa?ic? oa ia ai?iiiiai?iieeo SourceQ.
Aeenioe?aoe?th eiiiiiaio?a eiiieaeno i?ae ae??th NaSCN oa iaiaaeaiee
i?ioaie?c oeeo eiiiiiaio?a i?iaiaeeee, ye iienaii (Bec, 1989). Aieea
EF-1 ia eaoae?oe/io aeoeai?noue ai?iiaoeee-o?IE neioaoac aea/aee a 100
iee noaiaea?oii? noi?o? aeey ai?iiaoeeethaaiiy o?IE (Bec et al, 1989) c
aeiaeaaaiiyi 10% ae?oea?eio oa 100 ieI GTP aai GDP. Aei noi?o? aiineee 5
— 50 iiieue EF-1 aai eiio?ieueieo a?ee?a. Aieea EF-1 ia ATP/PPi iai?i,
yeee eaoae?co? oai?eaeai?e-o?IE neioaoaca, aea/aee a 100 iee noi?o?, ui
i?noeea 100 iI IA?AS ?I 7.5, 5 iI IgNl2, 1 iI oai?eaeai?i, 2.5 iI AO?,
1iI 32?-??? (100 E?/iieue), 1 ia/ie ANA, 0.5 iI ae?o?io?a?oie, 10 %
ae?oea?ei.

Ia?iaaa?e?caoe?th noniaic?? ee?oei NII cae?enithaaee 75 iea/ie
naiii?iii, ee?oei VH 25 in situ – 125 iea/ie naiii?iii a iioei?ciaaieo
aeey ??cieo ee?oei oiiaao. A?ineioac a?eea a ia?iaaa?e?ciaaieo ee?oeiao
VH25 aea/aee i?ae /an ?o ?ieoaaoe?? a noi?o?, ui i?noeea 130 iI
oeoe?ico, 50 iI aoeaoao eae?th, 50 iI KCl, 20 iI HEPES, pH 7.4, 0.5 iI
DTT, 5 iI ATP, 13 iI oinoie?aaoei, 5 iI aetheico, 6 iI aoeaoao iaai?th,
0.4 iI GTP, 30 iea/ie e?aaoeie?iaco, 240 ieM ?aae?iaeoeaio ai?iieeneioo
oa ?aooo iai?/aieo ai?iieeneio, 240 ieM eiaeii?, i?e 370C. Aeey ee?oei
NII e?ioeaa? eiioeaio?aoe?? aeayeeo ?aaaaio?a aoee ?ioeie, cie?aia: 0.8
iI ATP, 0.07 iI GTP, 7.5 iI oinoie?aaoei, 1.3 iI aoeaoao iaai?th, noi?o
i’yoe /e i’yoiaaeoeyoe 14N-ai?iieeneio oa ?aooo iai?/aieo ai?iieeneio,
250 ieI eiaeia. ?aae?iaeoeai?noue, ui caeeoaeanue a OOI-inaae? i?ney
eei’yo?iiy, na?ae/eea i?i e?euee?noue neioaciaaieo iie?iaioeae?a, a
??cieoey i?ae ?aae?iaeoeai?noth inaae?a, ui aoee io?eiai? ia?iaeith
aa?y/ith oa oieiaeiith OOI, aeacoaaea ia e?euee?noue ai?iiaoeee-o?IE a
?ieoaaoe?ei?e noi?o?. O?aineyoe?th iie?(U) oa aeia?iiai? i?IE a
a?eieneioacoth/?e nenoai? ?c ca?iaee?a ioaieoe? oa aiaeiaaiiee a?eeiaee
neioac a e?cao? ?aoeeoeioeeo?a e?iey i?iaiaeeee ye iienaii (Eeaiain,
1987).

?IEaca A c i?aeoeoieiai? caeice aeea oa o?IE aoee iiaeeo?eiaai?
a?aeiia?aeii O?OOe oa oeoi?anoea?inai?ea?aaceaeii ca?aeii ?ai?oa
iienaieo iaoiaeee (Melan et al, 1992). C iaoith oeoi?anoeaioii?
iiaeeo?eaoe?? EF-1 oa EF-1 ae?ae?coaaee i?ioe 100 iI ea?aiiaoiiai
aooa?a, pH 8. 3, 50 iI NaCl, 20% ae?oea?eio oa ?ieoaoaaee a i?enooiino?
O?OOe (0.1 ia/ie). I?/aiiy coieiyee NH4Cl (e?ioeaaa eiioeaio?aoe?y 50
iI), ? a?eee a?aeie?aiethaaee a?ae O?OOe o?iiaoia?ao??th ia naoaaeaen?
G-25. Aioo??oiueiee?oeiio oethi?anoeaioe?th aea/aee ia aeeaeo aai
o?eniaaieo 3.7% oi?iaeueaeaa?aeii ee?oeiao, aeei?enoiaoth/e
eiioieaeueiee i?e?ineii Bio-Rad MRC-600 aai na?oeiaee i?e?ineii,
iaeaaeiaiee aeey ?a?no?aoe?? oeoi?anoeaioe??.

Oeoi?anoeaioe?th o?eioioai?eiaeo caeeoe?a a iieaeoe? EF-1 aei??thaaee ia
niaeo?ioeoi?eiao?? SLM-8100 (SLM/Aminco) i?e aeiaaeei? oaee? caoaeaeaiiy
297 ii ?c oe?eiith u?eeie 2 ii aeey caoaeaeoth/iai i?iiaiy ? 8 ii aeey
i?iiaiy, ui aeionea?oueny. Ocaaaeueiai? niaeo?e ai?n?? aoee io?eiai?
aeey 10-30 aei??thaaiue a?ae 310 aei 500 ii c ?ioa?aaeii a 1 ii.
Iaiaaeaiee ?IEaciee a?ae?ie?c oa iiaeeo?eaoe?th o?IE
aoeei?o?icina/iaeiith i?iaiaeeee, ye iienaii ?ai?oa (Boutorin et al.,
1981, Jekowsky et al., 1977, Iao?ooaiei e ae?, 1990). Ooai?aiiy
/aoaa?oeiiiai eiiieaeno EF-1-GDP-o?IEPhe-oai?eaeai?e-o?IE neioaoaca
ia?aa??yee a 0.7% aaa?iciiio aae?, niinoa??aath/e ca cao?eieith
32?-o?IEPhe a aae? aiane?aeie aca?iiae?? c a?eeoi.

Eiiia?oiaioae?caoe?y oa /aiae?ia ai?iiaoeee-o?IE a ee?oeiao nnaaoe?a. Ia
ia?oiio aoai? aeine?aeaeaiue ooieoe?iiaeueii? eiiia?oiaioae?caoe?? ie
aaaaeaee ca iaiao?aeia aeiaanoe i?eioeeiiao iiaeeea?noue /aiae?iaa
ai?iiaoeee-o?IE aaciina?aaeiuei a ee?oeiao nnaaoe?a. Aeey aeinyaiaiiy
oe??? iaoe iio??aii aoei ?ic?iaeoe niin?a aeinoaaee ai?iiaoeee-o?IE aei
oeeoiieacie, ine?eueee ca ii?iaeueieo oiia o?IE ia caeaoia i?ioiaeeoe
e??cue ee?oeiio iaia?aio. I?e oeueiio aoei aaniethoii iaiao?aeii
aeio?eioaaoeny aeaio oiia: caaacia/eoe a?eueiee aeinooi aeciaaiieo
iae?iiieaeoe aei ee?oeie oa iaeneiaeueii caa?aaoe i?e oeueiio
?ioaeoi?noue i?aai?caoe?? ?? a?eie-neioacoth/iai aia?aoo.

Aeey ia?oi?aoe?? oeeoiieaciaoe/ii? iaia?aie ee?oei NII aoei ia?aii aeaa
iaoiaee – aeaeo?iii?aoe?th oa ia?iaeo aeaoa?aaioaie, cie?aia naiii?iii.
Ia?iaaa?e?caoe?y naiii?iii aeyaeeany a?eueo i’yeith i?ioeaaeo?ith, yea
ia ooeiaeaeoaaea aioo??oiueiee?oeiio i?aai?caoe?th aia?aoo a?eeiaiai
neioaco. A oaaeeoe? 1 iaaaaeaii aeai? oeiiaiai aenia?eiaioo ii
aecia/aiith ?iciiae?eo ?IE oa a?eea a ee?oeiao NII, ui ?ieoaoaaeeny a
i?enooiino? 3I-o?eaeeio aai noi?o? 3I-ai?iieeneio, i?ae
ia?iaaa?e?ciaaieie ee?oeiaie oa i?aeee?oeiiei i?inoi?ii. C oaaeeoe? 1
aeaeii, ui ia?iaea naiii?iii i?ecaiaeeoue aei iacia/iiai ciaioaiiy

Oaaeeoey 1.

Aeo?ae i?aneioaciaaieo 3I-?IE oa 3I-a?eea c ia?iaaa?e?ciaaieo ee?oei
NII.

O?aeoe?yE?euee?noue ee?oei, %?IE, %E?euee?noue ee?oei, %A?eie, %

?ioaeoi? ee?oeie

100

100

100

100Noia?iaoaio i?ney ia?iaaa?e?caoe??

_

13

_

19Ee?oeie i?ney ia?iaaa?e?caoe??

78

90

-Noia?iaoaio i?ney i?iieaaiiy ee?oei_6_8Ee?oeie i?ney i?iieaaiiy

81

81

84

66

e?eueeino? ?ioaeoieo ee?oei, i?e/iio e?euee?noue ?IE oa a?ee?a, ui
aea?eueieeenue ?c ia?iaaa?e?ciaaieo ee?oei, i?eaeecii aei??aith?
e?eueeino? c?oeiiaaieo ee?oei. Oaeei /eiii, aeayea iiioeyoe?y ee?oei
(aeecueei 20%) ? /ooeeaith aei naiii?io oa ?oeio?oueny i?ae /an
ia?iaaa?e?caoe??, aea?eueiyth/e naith ?IE oa a?eie iaciai?, a
noia?iaoaio, aea ?aooa ee?oei, iacaaaeath/e ia ia?oi?iaai?noue ?o
iaia?ai, caa??aathoue ?IE oa a?eie. Ne?ae i?aee?aneeoe, ui aioo??oi?e
ia’?i oaeeo ee?oei ? oe?eeii aeinooiiei aeciaaiiei iae?iiieaeoeai, ye?
oaeaeei oa ??aiii??ii ?iciiae?eyeeny a oeeoiieaci? ia?iaaa?e?ciaaieo
ee?oei. Inoaii? aoei aeiaaaeaii eiioieaeueiith i?e?ineii??th ee?oei, aei
yeeo aoei aeiaeaii oeoi?anoeaioii iiaeeo?eiaai? o?IE aai ?IEaco A.
Oaeei /eiii, io?eiai? aeai? na?ae/aoue i?i i?eioeeiiao iiaeeea?noue
eiiia?oiaioae?caoe?? ?IE oa a?ee?a a ee?oeiao nnaaoe?a. Aea iane?eueee
aeoeaieie a o?aineyoe?? i?IE ? ia?iaaa?e?ciaai? ca cai?iiiiiaaiith
iaoiaeeeith ee?oeie? Aeey a?aeiia?ae? iaiao?aeii ii??aiyoe oaeaee?noue
a?eeiaiai neioaco a ia?iaaa?e?ciaaieo oa ?ioaeoieo ee?oeiao. I?ney
iioei?caoe?? eiioeaio?aoe?e iecueeiiieaeoey?ieo eiiiiiaio?a
a?eie-neioacoth/iai aia?aoo a ia?iaaa?e?ciaaieo ee?oeiao aoei aeinyaiooi
i?aeoe/ii ?aeaioe/io c ?ioaeoieie ee?oeiaie oaeaee?noue o?aineyoe?? ia
i?ioyc? uiiaeiaio 30 oaeeei (?en. 1). Oaeei /eiii, aia?oa aoei noai?aii
nenoaio niieo/aii? ia?iaaa?e?caoe??-o?aineyoe?? a ee?oeiao nnaaoe?a, yeo
iiaeia aeei?enoiaoaaoe aeey aea/aiiy no?oeoo?ii-ooieoe?iiaeueii?
i?aai?caoe?? aia?aoo a?eeiaiai neioaco in vivo. Aeinyaiaiiy aeniei?
aoaeoeaiino? a?eeiaiai neioaco na?ae/eoue i?i iaeneiaeueia caa?aaeaiiy
?ioaeoiino? aia?aoo o?aineyoe?? a ia?iaaa?e?ciaaieo ee?oeiao. Oea oaeiae
i?aeoaa?aeaeo? iecueeee ??aaiue aeoiaeo c ieo iae?iiieaeoe a?eeiai? oa
?eaiioeea?iiai? i?e?iaee (aeea. oaae. 1). Aea a?aee?eoei caeeoaeiny
ieoaiiy, /e caeaoi? iecueeiiieaeoey?i? eioaeoi?e o?aineyoe?? a?eueii
aeenioe?thaaoe ?c ia?oi?iaaieo ee?oei? Aeey ia?aa??ee iiaeeeaino?
eiiia?oiaioae?caoe?? iecueeiiieaeoey?ieo niieoe, ye? aa?ooue o/anoue o
o?aineyoe??, aoei aea/aii ?o aieea ia aoaeoeai?noue a?ineioaco a?eea a
ia?iaaa?e?ciaaieo ee?oeiao NII. Aeai? oeiiaiai aenia?eiaioo iaaaaeaii a
oaae. 2. Aeaeii, ui aeeo/aiiy AO? aai AO? ?acii c GTP

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40000002e011800030000001e0017000000fb029a000000000000009001000000cc03020
11254696d6573204e657720526f6d616e2043797200cc65040000002d010600050000000
9020000000012000000320ab6091d0607000000d7e0f12c20f5e200660045003c0025002
20045004500040000002d01040004000000f0010600040000002701ffff040000002e010
100040000000201020005000000090200000000040000002d01040004000000f00103000
400000034020200030000003500030000000000

?en. 1. Aeeth/aiiy 14N-ai?iieeneio aei OOI-ia?ic/eiiiai inaaeo a
ia?iaaa?e?ciaaieo naiii?iii (1) aai ?ioaeoieo (2) ee?oeiao NII.
Iaaaaeaii aeeth/aiiy i?oee a 9.4*107 ?ioaeoieo aai ia?iaaa?e?ciaaieo
ee?oei.

Oaaeeoey 2.

Aaaeeea?noue iecueeiiieaeoey?ieo niieoe oa e?aaoeie?iace aeey a?eeiaiai
neioaco a ia?iaaa?e?ciaaieo ee?oeiao NII.

Aeeo/ai? c noi?o? aeey a?eeiaiai neioaco eiiiiiaioeAeeth/aiiy
14N-oai?eaeai?io aei neioaciaaieo iie?iaioeae?a, %iiaia
noi?o100AOP54GTP90AOP e GTP49Oinoie?aaoei4E?aaoeie?iaca89Noi?o
ai?iieeneio56Mg2+5K+(aeiaeaii Na+)13Aeeo?io?a?oie85

?c noi?o? aeey a?eeiaiai neioaco, a ye?e i?iaiaeeee ?ieoaaoe?th ee?oei,
ciaiooaaei oaeaee?noue a?eeiaiai neioaco eeoa iaiieiaeio. A?aenooi?noue
aeciaaiiiai GTP cian?i ia aieeaaea ia o?aineyoe?th. Ia niinoa??aaeinue
cia/iiai aoaeoo ? i?e a?aenooiino? aeciaaiii? e?aaoeie?iace.
Ea?aeeiaeueiee iiceoeaiee aieea ia a?eeiaee neioac iaea eeoa
i?enooi?noue aeciaaiiiai oinoie?aaoeio. Iiaeia c?iaeoe aeniiaie, ui any
AO?, ui caeeoa?oueny a ee?oeiao i?ney ia?iaaa?e?caoe??, oaeaeei
ooee?co?oueny, aea iiaea aooe aoaeoeaii ?aaaia?iaaia a i?enooiino?
aeciaaiiiai oinoie?aaoeio. A?aenooi?noue ?noioii? caeaaeiino? a?ae
aeciaaiieo oinoie?aaoeie?iace oa GTP iiaea na?ae/eoe i?i aoiaeaeaiiy aei
o?aineyoe?eiiai eiiia?oiaioo oa?iaio?a, ye? ia iathoue i?yiiai
a?aeiioaiiy aei o?aineyoe?? – e?aaoeie?iace oa ioeeaiceaeaeeoinoie?iace.
Oe?eaai, ui ca a?aenooiino? aeciaaiieo ai?iieeneio oa a oiiaao cia/iiai
?icaaaeaiiy ee?oeiiiai ai?noo a ia?iaaa?e?ciaaieo ee?oeiao oaeaee?noue
a?eeiaiai neioaco iaaeaea o?eueee oaea?/?. Oea na?ae/eoue ia ei?enoue
eiiia?oiaioae?caoe?? ai?iieeneio o ee?oeiao nnaaoe?a [Smith et al.,
1988, Samarel et al., 1991]. Oaeiae oe?eaaei ? io?eiaiee ?acoeueoao, ui
aeiaeaaaiiy iaaeeeoeo o?IE (aei 1 ia/ie) aei ia?iaaa?e?ciaaiieo ee?oei
i?ye ia aieeaa? ia a?eeiaee neioac. Oe? aeai? oa a?aenooi?noue
aeenioe?aoe?? ?IE c ia?iaaa?e?ciaaieo ee?oei (aeea oaae.1) aoee ia?oeie,
ui na?ae/aoue i?i iiaeeeaoeiiia?oiaioae?caoe?th o?IE a ee?oeiao
nnaaoe?a. Aeyaeaia iaie a?aenooi?noue ?noioii? ??cieoe? a niaeo?ao
a?ee?a, ui aoee neioaciaai? a ia?iaaa?e?ciaaieo oa ?ioaeoieo ee?oeiao,
na?ae/eoue, ui i?IE oaae iiaeooue caeeoaoeny ?iia?e?ciaaieie a
ia?oi?iaaieo ee?oeiao. Oaeei /eiii, nenoaia a?eeiaiai neioaco a
ia?iaaa?e?ciaaieo ee?oeiao ye e?euee?nii, oae ? ye?nii a?aeiia?aea?
a?eeiaiio neioaco a ?ioaeoieo ee?oeiao. Oiio oaea nenoaia iiaea aooe
aeei?enoaia aeey ae??oaiiy iaeiiai c iniiaieo caaaeaiue ?iaioe — aeieaco
eiiia?oiaioae?caoe?? oa /aiae?iaa ai?iiaoeee-o?IE in vivo.

sseui ai?iiaoeee-o?IE ae?enii eiiia?oiaioae?ciaaia a ee?oeiao NII, aiia
ia iiaeiia aeoiaeeoe ?c ee?oei ? i?ney ?o ia?iaaa?e?caoe??. Oiio
aea/ath/e ?iciiae?e ai?iiaoeee-o?IE i?ae aioo??oiueiee?oeiiei oa
iicaee?oeiiei ia’?iaie, iiaeia i?aeoaa?aeeoe /e ni?inooaaoe ?nioaaiiy
oaei? eiiia?oiaioae?caoe??. Ioaea, noi?o 15 14N-ai?iieeneio oa
a?aeiia?aeieo 3I-ai?iiaoeee-o?IE aoea aeiaeaia aei ia?iaaa?e?ciaaieo
ee?oei. Oeaio?eooaoaaiiyi ae?eaio ee?oei /a?ac iaai? /ania? ?ioa?aaee
aoei aecia/aii a?aeiinio e?euee?noue aeciaaiii? (3I) ai?iiaoeee-o?IE,
yea neoaoaaea ca aioo??oi?e eiio?ieue, oa aiaeiaaiii neioaciaaii? (14N)
ai?iiaoeee-o?IE o ee?oeii?e o?aeoe?? oa a noia?iaoaio? (?en.2).
Aeyaeeiny, ui ?iciiae?e aiaeiaaiii? oa aeciaaiii? ai?iiaoeee-o?IE i?ae
oeeie aeaiia o?aeoe?yie aeoaea a?ae??ciy?oueny. Aeey aeciaaiii?
ai?iiaoeee-o?IE ni?aa?aeiioaiiy ?? e?eueeino? ona?aaeei? oa iica
ee?oeiaie aoei aea?aaeaioia ni?aa?aeiioaiith ee?oeiiiai oa
iicaee?oeiiiai ia’?i?a (i?eaeecii 0.2), ui na?ae/eei i?i aacia?aoeiaeiee
iai?i aeciaaiii? ai?iiaoeee-o?IE i?ae iicaee?oeiiei oa
aioo??oiueiee?oeiiei i?inoi?aie. Iaaiaee, o aeiaaeeo aiaeiaaiii?
ai?iiaoeee-o?IE niinoa??aaeany ?? cao?eiea (a iaaaaeaiiio aenia?eiaio?
a?eueo i?ae a 25 ?ac?a a ii??aiyii? c aeciaaiiith ai?iiaoeee-o?IE) oa
ianooiia aeoaea iia?eueia aeenioe?aoe?y ?c ia?iaaa?e?ciaaieo ee?oei.

Ua iaeiei e?eoa???i eiiia?oiaioae?caoe?? aiaeiaaiii? ai?iiaoeee-o?IE
iiaea aooe ?? no?ee?noue aei ae?? aeiaeaii? aei ia?iaaa?e?ciaaieo ee?oei
?IEace. Noaia aenia?eiaioo aoea oaeith ae — 3I-ai?iiaoeee-o?IE
aeiaeaaaee aeciaaiii, a 14N-ai?iiaoeee-o?IE neioacoaaeany
aaciina?aaeiuei a ee?oeiao. Aieea ?IEace A ia aeciaaiio oa aiaeiaaiio
ai?iiaoeee-o?IE iaaaaeaii ia ?en.3. Aeaeii, ui 3I-?aae?iaeoeai?noue
oaeaeei cieea? i?e ?IEaci?e ia?iaoe?,

?en.2 Ni?aa?aeiioaiiy aioo??oiueiee?oeiii? oa iicaee?oeiii?
ai?iiaoeee-o?IE i?ney ?ieoaaoe?? ia?iaaa?e?ciaaieo ee?oei NII ?c
noi?ooth 14N-ai?iieeneio (1) oa a?aeiia?aeieo 3I-ai?iiaoeee-o?IE (2). 3
– ni?aa?aeiioaiiy ee?oeiiiai oa ciai?oiueiai ia’?i?a a aeaiiio
aenia?eiaio?.

?en.3. Aieea ?IEace A ia aiaeiaaiio (1) oa aeciaaiio (2) ai?iiaoeee-o?IE
a ia?iaaa?e?ciaaieo ee?oeiao NII, a oaeiae ia aiaeiaaiio ai?iiaoeee-o?IE
(3) a aiiiaaiao? ee?oei. 100% aeey eiaeiiai aeiaaeea — e?euee?noue
ai?iiaoeee-o?IE ca a?aenooiino? ?IEace.

aeacoth/e ia iiaio aeinooii?noue aeey ?IEace aeciaaiii? ai?iiaoeee-?IE.
A oie aea /an aiaeiaaiia 14N-ai?iiaoeee-o?IE caoeuaia a?ae ae?? ?IEace.
Ne?ae i?aee?aneeoe, ui iecueea aoaeoeai?noue ?IEaciiai a?ae?ie?co
14N-ai?iiaoeee-o?IEia ? iane?aeeii oo?oaeiaiiai aeinooio ?IEace A aei
ia?iaaa?e?ciaaieo ee?oei, ai, ye caaaeoaaeiny aeua, oeoi?anoeaioii
iiaeeo?eiaaia ?IEaca A ia? iaiaiaaeaiee aeinooi aei oaeeo ee?oei ?
?iciiae?ey?oueny ona?aaeei? ee?oeie aeineoue ??aiii??ii. A?eueo oiai,
yeui ia?iaaa?e?ciaai? ee?oeie, ui i?noyoue aiaeiaaiio oa aeciaaiio
ai?iiaoeee-o?IE, aiiiaai?coaaoe ia?aae ia?iaeith ioeeaacith, oi ia
niinoa??aa?oueny i?yei? ??cieoe? o ??ai? a?ae?ie?co iaio aeae?a
ai?iiaoeee-o?IE (?en.3, e?eaa 3).

Oaeei /eiii, a iienaieo aeua aenia?eiaioao ie iaea?aeaee ia?o? nooo?a?
aeieace eiiia?oiaioae?caoe?? ai?iiaoeee-o?IE a ee?oeiao nnaaoe?a. Aea
caeeoaeinue iaa?aeiiei, /e ?nio? ca’ycie i?ae eiiia?oiaioae?caoe??th
aia?aoo o?aineyoe?? oa aoaeoeai?noth a?eeiaiai neioaco. ?iaeoa eaaeo/e,
iio??aii aoei aecia/eoe iiaeeeaa ooieoe?iiaeueia cia/aiiy
eiiia?oiaioae?caoe?? ai?iiaoeee-o?IE. Na?aae iioaioe?eieo iaoai?ci?a,
ui ?aae?cothoue ia?aaaao eiiia?oiaioae?caoe??, /?eueia i?noea caeia?
/aiae?ia, aai aaciina?aaei?e, aac aeoiaeo a ?ic/ei, o?ainii?o i?ii?aeieo
i?iaeoeo?a i?ae iine?aeiaieie o/anieeaie eiie?aoiiai iaoaaie?/iiai oeyoo
(Srere, 1987, Nie?ei, 1989). Aieiaiei e?eoa???i ?nioaaiiy /aiae?iaa
iaoaaie?o?a ? iaiiaeeea?noue caeo/aiiy aei iaoaaie?/iiai eaioethaa
i?aneioaciaaieo aeciaaiii i?ii?aeieo i?iaeoeo?a oeeo ?aaeoe?e, oiio ui,
ca?aeii aecia/aiith, eaiae?ciaaiee i?ioeann ? cae?eoei aei inoaii?o
(Srivastava et al, 1987). Ine?eueee i?ii?aeia niieoea a?ineioaco a?eea –
ai?iiaoeee-o?IE, oi, ii??aiyaoe ??aaiue aeei?enoaiiy aeey a?eeiaiai
neioaco aiaeiaaiii? oa aeciaaiii? ai?iiaoeee-o?IE, iiaeia c?iaeoe
aeniiaie a?aeiinii /aiae?iaa oeueiai iaoaaie?oo a ee?oeiao nnaaoe?a.
Ia?aaaaeia aeei?enoaiiy aiaeiaaiii?, neioaciaaii? aaciina?aaeiuei a
ee?oeiao ai?iiaoeee-o?IE aoei a i?yiei iiceoeaiei na?ae/aiiyi uiaei
?nioaaiiy iaoai?cio /aiae?iaa a a?eeiaiio neioac?.

Aeey ia?aa??ee oeueiai iieiaeaiiy ciia aoee aeei?enoai? aenia?eiaioe ?c
iiaea?eiei i?/aiiyi, a yeeo aiaeiaaiii neioaciaaia ai?iiaoeee-o?IE aoea
14N-i?/aiith, a i?aneioaciaaia aeciaaiia ai?iiaoeee-o?IE — 3I-i?/aiith.
A ee?oeiiiio aeno?aeo? ia aoei aeyaeaii ia?aaaae aeiaeii?
ai?iiaoeee-o?IE o aeei?enoaii? aeey a?eeiaiai neioaco. Oea na?ae/eei i?i
a?aenooi?noue /aiae?iaa aiaeiaaiii? ai?iiaoeee-o?IE a aacee?oeii?e
a?eie-neioacoth/ee nenoai?. Aea iaiiaeeea?noue i?iaeaiiino?oaaoe
/aiae?ia in vitro ua ia na?ae/eoue i?i a?aenooi?noue oeueiai iaoai?cio
aaciina?aaeiuei a ee?oeiao. Oe?eeii iiaeeeai, ui aeey ooieoe?iioaaiiy
iaoai?cio /aiae?iaa iaiao?aeia ?ioaeoia i?aai?caoe?y a?eie-neioacoth/iai
aia?aoo, yeo iiaea aooe c?oeiiaaii i?ae /an aiiiaai?caoe?? ee?oei. sse
iieacaii aeua, ?ic?iaeaia iaie nenoaia ia?iaaa?e?caoe??-o?aineyoe??
caeeoa? aia?ao a?eeiaiai neioaco iaeaea iaaeioi?eaiei. Aa?o/e oea aei
oaaae, aiaeia?/i? aenia?eiaioe aoei i?iaaaeaii a ia?iaaa?e?ciaaieo
ee?oeiao. 3I-ai?iiaoeee-o?IE oa 14N-ai?iieeneioe aoei aeiaaaeaii
aaciina?aaeiuei aei ia?iaaa?e?ciaaieo ee?oei, ? aeeth/aiiy eiaeii? i?oee
aei iie?iaioeaeiiai eaioethaa ii??aithaaee ?c ??aiai a?aeiia?aeii?
ai?iiaoeee-o?IE a ee?oeiao ia oie aea /an. Ni?aa?aeiioaiiy oeeo
ia?aiao??a aoei iieacieeii aoaeoeaiino? aeei?enoaiiy aeciaaiii? oa
aiaeiaaiii? ai?iiaoeee-o?IE. sse aeaeii c ?en.4, iieacieee aoaeoeaiino?
aeei?enoaiiy ai?iiaoeee-o?IE aeoaea a?ae??ciythoueny aeey aiaeiaaiiiai
oa aeciaaiiiai iiia?aaeiueiai i?iaeoeo?a. 14N-?cioii ooee?co?oueny a 15
?ac?a a?eueo aoaeoeaii i?ae 3I-?cioii, oiaoi aiaeiaaiia ai?iiaoeee-o?IE
? iaaaaaoi a?eueo aoaeoeaiei i?ii?aeiei i?iaeoeoii aeey a?ineioaco
a?eea, i?ae aeciaaiia. Aiaeia?/i? ?acoeueoaoe aoei io?eiaii a nenoai?
aeaeo?iii?iaaieo ee?oei NII. Iaiao?aeii caoaaaeeoe, ui ia?aaaaeia
aeei?enoaiiy aiaeiaaiii? ai?iiaoeee-o?IE niinoa??aaeiny iacaaaeath/e ia
oa, ui, ca?aeii c aeaieie eiioieaeueii? i?e?ineii??, aeciaaiia

?en. 4. A?eeiaee neioac a ia?iaaa?e?ciaaieo naiii?iii ee?oeiao NII c
aeei?enoaiiyi aiaeiaaiieo (A) oa aeciaaiieo (A) ai?iiaoeee-o?IE. 1 –
aeeth/aiiy i?oee aei ai?iiaoeee-o?IE, 2 – aeeth/aiiy i?oee aei
iie?iaioeaeieo eaioetha?a. E?euee?noue ee?oei a aeaiiio aenia?eiaio? –
3.8*106.

o?IE oaeaeei iio?aiey? aei ia?iaaa?e?ciaaieo ee?oei ? ?iciiae?ey?oueny a
ieo aeineoue ??aiii??ii.

Ioaea, iieacaii, ui iaeaeaa e?eoa??y /aiae?iaa — oo?oaeiai?noue
aeei?enoaiiy aeciaaiiiai i?ii?aeiiai i?iaeoeoo, oae naii ye ? no?oeoo?ia
a?aeie?aieai?noue aiaeiaaiiiai iiia?aaeieea, aeeiiothoueny i?ae /an
a?ineioaco a?eea a ee?oeiao nnaaoe?a. Oaeei /eiii, io?eiaii ia?o?
aenia?eiaioaeuei? aeai?, ye? na?ae/aoue i?i ?nioaaiiy ooieoe?iiaeueii?
eiiia?oiaioae?caoe?? ai?iiaoeee-o?IE. *aiae?ia aiaeiaaiii?
ai?iiaoeee-o?IE i?ecaiaeeoue aei aoaeoeaiiai, ia a?aei?io a?ae
aeciaaiii? ai?iiaoeee-o?IE, ?? aeei?enoaiiy. Ia aac? aeaieo, ui
aeeeaaeaii a aeaiiio ?icae?e?, oa aa?o/? aei oaaae a?aeiio aei ii/aoeo
iaoeo aeine?aeaeaiue noaio oeeeea aeiiaaoe?? noaei iiaeeeaei oyaeoe
a?iioaoe/iee oeeee /aiae?iaa ai?iiaoeee-o?IE oa o?IE i?ae /an
o?aineyoe?? (?en.5). O oaeiio oeeee? ai?iiaoeee-o?IE ia?aaea?oueny a?ae
ai?iiaoeee-o?IE neioaoace aei oaeoi?a aeiiaaoe?? EF-1 ? aeae? ia
?eainiio aac aeoiaeo aei ?ic/eia. Ine?eueee aenia?eiaioe ia ee?oeiiiio
??ai? ia aeicaieyee noaeeoe i?i iiaeeeaiai iina?aaeieea a ia?aaea/? o?IE
a?ae ?eainiie aei ai?iiaoeee-o?IE neioaoace, oiiaii aoei iicia/aii, ui
o?IE ia?aaea?oueny c ?eainiie i?yii aei ai?iiaoeee-o?IE neioaoace.
Cai?iiiiiaaii, ui /aiae?ia ai?iiaoeee-o?IE a?ae a?eea aei a?eea
a?aeaoaa?oueny a niaoe?aeueieo o?aineyoe?eieo eiiia?oiaioao,
i?aai?caoe?y yeeo ?oeio?oueny i?e aiiiaai?caoe??.

Eieae?caoe?y oaeoi?a aeiiaaoe?? 1 a o?a?iaeanoao ethaeeie. Aoaeo
ooieoe?iiaeueii? aeoeaiino? aia?aoo o?aineyoe??. Ianooiiei caaaeaiiyi
aoei io?eiaoe a?eueo aeieeaaeio ?ioi?iaoe?th i?i ?nioaaiiy
o?aineyoe?eieo eiiia?oiaio?a oa iiaeeea?noue ci?i a ?o i?aai?caoe?? ca
??cieo oiia ooieoe?iioaaiiy ee?oeie. Aoei c?iaeaii

?en.5 A?iioaoe/ia noaia /aiae?iaa ai?iiaoeee-o?IE i?ae /an aeiiaaoe??
aoea??ioe/iiai a?eeiaiai neioaco. A?N – ai?iiaoeee-o?IE neioaoaca, A, ?,
oa A – naeoe ca’ycoaaiiy o?IE ia ?eainii?, /i?i? e?oae iicia/athoue
ai?iiaoeee-o?IE, n??? e?oae – iaioeaeee- aai aeaaoeeeueiaaio o?IE

i?eiouaiiy, ui i?iaeaiiino?oaaoe iioaioe?eio eaa?euei?noue
o?aineyoe?eieo eiiia?oiaio?a iiaeeeai ca ??cieo aia?aaoe/ieo noai?a

ee?oeie, ui noi?iaiaeaeothoueny ci?iaie o?aineyoe?eii? aeoeaiino?. sse
ia?ea?e aoei aeei?enoaii a?eee, ui ? iaea?eueo eiia??ieie eiiiiiaioaie
oaeeo eiiia?oiaio?a — ? noaiaeeieoe? oaeoi?a aeiiaaoe?? EF-1.
Caaaeaiiyi aeaiiai aoaio aeine?aeaeaiue aoei c’ynoaaoe, /e ci?ith?oueny,
? yeei /eiii, aioo??oiueiee?oeiia eieae?caoe?y EF-1 e EF-1 a caeaaeiino?
a?ae ai?noo oinoie?aaoeio, a?aenooi?noue yeiai a ?ieoaaoe?ei?e noi?o?,
ye iieacaii iaie aeua, ni?e/eiy? iiaio coieieo a?eeiaiai neioaco a
ia?iaaa?e?ciaaieo ee?oeiao (aeea. oaae.2).

Ine?eueee aenia?eiaioe c aea/aiiy eieae?caoe?? a?ee?a e?aua i?iaiaeeoe
ia ?iia?e?ciaaieo ia iiaa?oi? ee?oeiao, ye iiaeaeue aoea aea?aia
ia?aeiia eoeueoo?a o?a?iaeano?a ethaeeie VH25, ye? ?inee i?ee??ieaieie
aei iiaa?oi? /aoie Iao??. I?ney aaeaioaoe?? iaoiaeo ia?iaaa?e?caoe??,
?ic?iaeaiiai iaie ?ai?oa aeey noniaic?? ee?oei NII, aei o?a?iaeano?a
ethaeeie, aoei iieacaii, ui oaeaee?noue o?aineyoe?? a ia?iaaa?e?ciaaieo
o?a?iaeanoao aoea e?i?eiith ia i?ioyc? 15-20 oaeeei. Oeueiai /ano aoei
oe?eeii aeinoaoiuei aeey i?iaaaeaiiy aenia?eiaio?a c aea/aiiy
eieae?caoe?? a?ee?a a i?enooiino? aai ca a?aenooiino? oinoie?aaoeio.

A oe?e ?iaio? canoiniaaii iaoiaee/iee i?aeo?ae aeey aea/aiiy
aioo??oiueiee?oeiii? eieae?caoe?? eiiiiiaio?a aia?aoo o?aineyoe??, yeee
aeaui a?ae??ciy?oueny a?ae noaiaea?oieo ?ioiioeoi?anoeaioieo
aeine?aeaeaiue. Aeei?enoaiiy ?ic?iaeaii? iaie nenoaie ia?iaaa?e?ciaaieo
ee?oei aea? ciiao eaaei iai?ioethaaoe a?eeiaeie eiiiiiaioaie aia?aoo
o?aineyoe??, oiaoi aeiaeaaaoe iiaeeo?eiaai? oeoi?anoea?i?cio?ioe?aiaoii
EF-1 oa EF-1 aaciina?aaeiuei aei ee?oei, ye? ciaoiaeyoueny a oiio /e
?ioiio ooieoe?iiaeueiiio noai?, oa aea/aoe aioo??oiueiee?oeiio
eieae?caoe?th oeeo a?ee?a aaciina?aaeiuei i?ae /an aeoeaii? o?aineyoe??,
aai eiee ?? coieiaii. Aeyaeaii, ui a ii?i? ?iciiae?e EF-1 a o?a?iaeanoao
ethaeeie ? a?eueo a?aioey?iei, i?ae ?iciiae?e oeueiai a?eeo a ee?oeiao,
?ieoaiaaieo aac oinoie?aaoeio (?en.6 a,a). Oe?eaaei aeyaeany oaeo
iayaiino? EF-1 a ee?oeiiiio yae??, iniaeeai ca a?aenooiino?
oinoie?aaoeio, eiee ia?aaaaeia a?eueo?noue iieaeoe oeueiai a?eeo ia?
yaea?io eieae?caoe?th. Ia a?aei?io a?ae EF-1, eieae?caoe?y EF-1 ia
caeaaeeoue a?ae i?enooiino? oinoie?aaoeio a ?ieoaaoe?ei?e noi?o? (?en.6
a,a). A iaio aeiaaeeao niinoa??aa?oueny /?oei ae?aaeaia a?aioey?ia
ea?oeia ?iciiae?eo oe??? noaiaeeieoe? oaeoi?a aeiiaaoe?? a oeeoiieaci?.
Oaeei /eiii, aeene?aoia oeeoiieaciaoe/ia eieae?caoe?y a i?enooiino?
oinoie?aaoeio aoea i?eoaiaiia iaii a?eeai, a oie /an eiee aeeeth/aiiy
oinoie?aaoeio ?c ?ieoaaoe?eii? noi?o? i?ecaiaeeei aei ia?a?iciiae?eo
EF-1, aea ia ci?ithaaei eieae?caoe?th EF-1.

Ne?ae aecia/eoe, ui aeey ciaioaiiy ianiaoe?o?/ieo aoaeo?a on?
aenia?eiaioe c oeoi?anoeaioii iiaeeo?eiaaieie noaiaeeieoeyie EF-1
i?iaiaeeeeny a i?enooiino? aeniei? eiioeaio?aoe?? (6 ia/ie)
iaaeueaoi?io. Aeey ia?aa??ee ooieoe?iiaeueiiai cia/aiiy ci?ie
eieae?caoe?? EF-1 aoei aea/aii ?iciiae?e a ee?oei? oeoi?anoeaioii
iiaeeo?eiaaiiai ?iaeoeaiaaiiai EF-1. Aeyaeaii, ui ?iaeoeaaoe?y EF-1
i?ecaiaeeea aei ciaioaiiy e?eueeino? oeueiai a?eea a oeeoiieaci? ? aei
iaa?iiaaeaeaiiy eiai o yae?? (?en.6ae), i?e/iio oey ea?oeia
niinoa??aaeany ca oiia ?ieoaaoe?? ee?oei o iiai?e, aeeth/ath/e
oinoie?aaoei, noi?o? aeey a?eeiaiai neioaco. Ioaea, aioo??oiueiee?oeiia
eieae?caoe?y iaaeoeaiiai EF-1 i?e aeoeaiiio a?eeiaiio neioac? aoea
iiae?aiith aei eieae?caoe?? aeoeaiiai EF-1 ca oiia, eiee
a?eie-neioacoth/ee aia?ao i?aeoe/ii ia ooieoe?iio?. Ia aac? oeeo aeaieo
iiaeia c?iaeoe i?eiouaiiy, ui eiee EF-1 ia i?eeia? o/ano? a o?aineyoe??,
a?eueoa /anoeia eiai iieaeoe ciaoiaeeoueny o ee?oeiiiio yae??. Ca oiia
iiaiiai caa?aaeaiiy ooieoe?iiaeueii? aeoeaiino? nenoaie o?aineyoe??,
eieae?caoe?y EF-1 oa EF-1 iiae?aia oa ia? aeaui a?aioey?iee oa?aeoa?.
O?aaa a?aecia/eoe, ui a?aioey?iee ?iciiae?e EF-1 o o?a?iaeanoao ethaeeie
aoa iauiaeaaii i?aeoaa?aeaeaiee ?ioiioeoi?anoeaioiei iaoiaeii (Sanders
et al., 1996). Ea?oeia aeene?aoii? eieae?caoe?? noaiaeeieoeue oaeoi?a
aeiiaaoe?? EF-1 ? iiae?aiith aei ?iciiae?eo a ie?aiaeaiae?ioeeoao ca?ai,
ye? i?noeee i?IE, ??IE, EF-1 oa a?a?i?e-o?IE neioaoaco (Barbarese et
al., 1995). Oae? ca?ia-a?aioee iiaeooue yaeyoe niaith o?aineyoe?ei?
eiiia?oiaioe, ye? ooieoe?iiothoue a oeeoiieaci? ia?aiaeo ee?oei. Iaie
anoaiiaeaii, ui iaaeoiaeaeaiiy oaeiai aaaeeeaiai eiiiiiaioo aia?aoo
o?aineyoe?? ye EF-1 aei cai?iiiiiaaieo o?aineyoe?eieo eiiia?oiaio?a
iiaea caeaaeaoe a?ae aoaeoeaiino? i?ioeano a?eeiaiai neioaco a ee?oei?
(?en.6). Oe? aeai? ?

?en.6 Eieae?caoe?y oeoi?anoeaioii iiaeeo?eiaaieo EF-1 (A,A), EF-1 (A,A)
oa ?iaeoeaiaaiiai EF-1 (Ae) a ia?iaaa?e?ciaaieo o?a?iaeanoao ethaeeie,
ui aoee ?ieoaiaai? a noi?o? aeey a?eeiaiai neioaco a i?enooiino?
(A,A,Ae) aai aac (A,A) oinoie?aaoeio.

ia?oeie, ui na?ae/aoue i?i iiaeeea?noue aia?aaoe/ii-caeaaeii? ci?ie
eieae?caoe?? eiiiiiaio?a aia?aoo o?aineyoe?? a aoea??ioe/i?e ee?oei?.

Aa?o/e ca iniiao iiaeaeue ooieoe?iioaaiiy eiiieaeno EF-1H (Janssen et
al, 1994) iiaeia ?ioa?i?aooaaoe iaea?aeai? iaie aeai? oaeei /eiii: EF-1
? /anoeiith noaoe?iia?iiai eiiieaeno EF- 1,annioe?eiaaiiai ?c ee?oeiieie
no?oeoo?aie, EF-1 ia?aaoaa? a oeueiio eiiieaen? eeoa i?ae /an GDP/GTP
iai?io ia eiai iieaeoe?. Aioo??oiueiee?oeiia eiioeaio?aoe?y EF-1
i?eiaeii? ia ii?yaeie ia?aaeuo? eiioeaio?aoe?th EF-1. Oiio i?e aeoeaiiio
a?eeiaiio neioac? niinoa??aa?oueny iiae?aia eieae?caoe?y EF-1 oa EF-1, a
i?e coieioe? o?aineyoe?? a?aeaoaa?oueny ci?ia eieae?caoe?? EF-1, aea ia
EF-1, yeee caeeoa?oueny ?iia?e?ciaaiei ia ee?oeiieo no?oeoo?ao.

Oe?eaaei ? oaeo eieae?caoe?? EF-1 a ee?oeiiiio yae??. Oe? aeai? aoee
iacaeaaeii i?aeoaa?aeaeai? iaoiaeaie ?ioiioeoi?anoeaioe?? oa
?ioiiaeaeo?iiii? i?e?ineii?? (Sanders et al, 1996, Barbarese et al.,
1995). ssaea?ia eieae?caoe?y EF-1 iiaea aooe i?iyaii aeeiiaiiy oe??th
iieaeoeith aeayeeo iaeaiii?/ieo ooieoe?e, cie?aia cai?iiiiiaaiith iaie
o/ano? EF-1 a o?ainii?o? uieii neioaciaaieo o?IE c yae?a aei
o?aineyoe?eieo eiiia?oiaio?a o oeeoiieaci?.

Ioaea, aia?oa anoaiiaeaii iiaeeea?noue ci?ie eieae?caoe?? eiiiiiaio?a
aia?aoo o?aineyoe?? o a?aeiia?aeue ia iaaaoeai? aieeae, cie?aia ia
aia?aaoe/io iaaeinoaoi?noue. Oe?eeii iiaeeeai, ui ia?a?iciiae?e iieaeoe
EF-1 a caeaaeiino? a?ae oinoie?aaoeio a?aeca?eaeth? caaaeuei? ci?ie a
no?oeoo?? eaa?eueieo, ia ca’ycaieo o?eaaei ?c ee?oeiieie no?oeoo?aie
eiiiiiaio?a o?aineyoe?eieo eiiia?oiaio?a. A?aenooi?noue ci?i
eieae?caoe?? EF-1 na?ae/eoue i?i oa, ui eiiieaen EF-1 iiaea aooe iaeiei
c aaciaeo eiiiiiaio?a oaeeo eiiia?oiaio?a, ye? caaacia/othoue
“i?ea’ycoaaiiy” eiiia?oiaio?a aei eiie?aoiiai i?noey neioaco oiai /e
?ioiai a?eea. E??i oiai, oe? ?acoeueoaoe na?ae/aoue ia ei?enoue
?nioaaiiy iiaiai ??aiy ?aaoeyoe?? a?eeiaiai neioaco, yeee iiaea
cae?enithaaoeny oeyoii anioe?aoe??/aeenioe?aoe?? o?aineyoe?eieo
eiiia?oiaio?a.

Iaeiei ?c eiiiiiaio?a oaei? ?aaoeyoi?ii? nenoaie iiaeooue aooe
a?ieia?/ii iaaeoeai? eiioi?ia?e o?IE, ye? c’yaeythoueny a i?aai?ci? ca
iani?eyoeeaeo oiia (Iaoeoea e ae?., 1972). ?ai?oa ie iieacaee, ui oae?
eiioi?ia?e caeaoi? ooai?thaaoe noaa?eueiee iai?iaeoeoeaiee eiiieaen c
ai?iiaoeee-o?IE neioaoacaie oa ?eainiiaie (Iaa?ooeeee e ae?., 1984,
Iaa?ooeeee e ae?., 1986), ? aenoioee i?eiouaiiy, ui oea iiaea iaoe iaaia
?aaoeyoi?ia cia/aiiy. A aeena?oaoe?? i?iaeaiiino?iaaii, ui i?enooi?noue
iaaeoeaieo eiioi?ia??a o?IE aieeaa? ia o?aineyoe?th i?e?iaeieo i?IE in
vitro, oiaoi iieacaii i?eioeeiiao iiaeeea?noue ?aaoeyoi?iiai aieeao
iaaeoeaieo eiioi?ia??a o?IE ia o?aineyoe?th. Oi?ioaaiiy noaa?eueieo
iai?iaeoeoeaieo eiiieaen?a o?IE c ?ioeie eiiiiiaioaie aia?aoo
o?aineyoe?? iiaea i?ecaiaeeoe aei ii?ooaiiy iaoai?cio ?o aoiaeo aei
o?aineyoe?eiiai eiiia?oiaioo aai iaa?oue aei ?iciaaeo oaeiai
eiiia?oiaioo. *e ae?enii oaeee iaoai?ci ooieoe?iio? o ee?oeiao nnaaoe?a,
ia? aooe ia?aa??aii a iiaeaeueoeo aeine?aeaeaiiyo.

Aca?iiae?y iae?iiieaeoe i?ae /an /aiae?iaa o?IE a i?ioean? o?aineyoe?? o
aoea??io. Oaeei /eiii, a aenia?eiaioao, i?iaaaeaieo c ee?oeiaie,
io?eiaii ?yae aeaieo, ui i?aeoaa?aeaeothoue iiaeeea?noue ?nioaaiiy a ieo
o?aineyoe?eieo eiiia?oiaio?a, ye? caaacia/othoue ooieoe?iioaaiiy
iaoai?cio /aiae?iaa oa iaiaaeothoue aeinooi ana?aaeeio aeciaaiieo
ai?iiaoeee-o?IE. E??i oiai, iieacaii, ui a?aenooi?noue oaeeo iae?ia?a?a
ye oinoie?aaoei a ee?oei? iiaea i?ecaiaeeoe aei ?iciaaeo eiiia?oiaio?a
/e, i?eiaeii?, aei aeoiaeo c ieo ie?aieo eiiiiiaio?a aia?aoo
o?aineyoe??. Ne?ae a?aecia/eoe, ui iaaeie?eii aenia?eiaio?a in vivo ?
oa, ui aiie caeaoi? ae??oeoe i?iaeaio eeoa a caaaeueiiio ieai?, oiaoi
i?aeoaa?aeeoe iiaeeea?noue /aiae?iaa ai?iiaoeee-o?IE a ee?oeiao, aea ia
aeathoue ?ioi?iaoe?th i?i aeaoaeueiee iaoai?ci /aiae?iaa. Oiio aoei
iaiao?aeiei ciiaeaethaaoe oe? aca?iiae?? a aeine?aeao in vitro,
aea/ath/e iiaeeea?noue anioe?aoe?? ?cieueiaaieo eiiiiiaio?a aia?aoo
o?aineyoe??, ui iathoue nii??aeiai?noue aei o?IE. Ine?eueee iniiaia
i?noea a eaioethc? /aiae?iaa caeia? a?iioaoe/ia ia?aianaiiy
ai?iiaoeee-o?IE a?ae ai?iiaoeee-o?IE neioaoace aei EF-1, ? i?yei?
?ioi?iaoe?? i?i anioe?aoe?th oeeo aeaio a?ee?a aei ii/aoeo iaoeo
aeine?aeaeaiue ia ?nioaaei, ie aaaaeaee ca aeioe?eueia c’ynoaaoe
i?eioeeiiao iiaeeea?noue oaei? aca?iiae?? a aeine?aeao in vitro, ia
??ai? ?iaeea?aeoaeueieo a?ee?a.

C oi/ee ci?o a?iioace /aiae?iaa iaea?eueo i?eaeaoiei aeey aea/aiiy
oeueiai iaoai?cio caea?oueny noaa?eueiee eiiieaen EF-1H c aae?e-o?IE
neioaoacith, yeee i?noeoue EF-1, a?eee, ui eaoae?cothoue GDP/GTP iai?i a
eiai iieaeoe?, oa ai?iiaoeee-o?IE neioaoaco. ?ai?oa aoei aeyaeaii, ui
e?iaoe/i? ia?aiao?e ?aaeoe?? ai?iiaoeeethaaiiy ia caeaaeaoue a?ae
anioe?aoe?? aae?e-o?IE neioaoace c EF-1H ? i?aeoe/ii iaeiaeia? aeey
a?eueii? oa ca’ycaii? c EF-1H neioaoace (Bec et al, 1989). I?ioa,
ca?aeii c a?iioacith /aiae?iaa, ui ia?aaeaa/a? o/anoue EF-1 a
aeenioe?aoe?? ai?iiaoeee-o?IE c ai?iiaoeee-o?IE neioaoace, oaea
aeenioe?aoe?y iiaeiia i?ioiaeeoe i?e oi?ioaaii? iio??eiiai eiiieaeno
aae?e-o?IE*EF-1*GTP. ?c oeueiai aeieeaa?, ui aeey aeyaeaiiy yeiainue
aoaeoo e?euee?noue iieaeoe EF-1 a ?ieoaaoe?ei?e noi?o? iiaeiia
aei??aithaaoe e?eueeino? iieaeoe i?iaeoeoa ?aaeoe??, ai?iiaoeee-o?IE, a
ia e?eueeino? iieaeoe oa?iaioo, ye niinoa??aa?oueny a eiiieaen?
aae?e-o?IE neioaoaca-EF-1H. Iaaeeeoie EF-1 iio??aai, uia caaacia/eoe
iiaeeea?noue iino?eiiai a?aeoieo ai?iiaoeee-o?IE a?ae aae?e-o?IE
neioaoace. Oaeaee?noue neioaco aae?e-o?IE a i?enooiino? iaaeeeoeo EF-1
oa GTP iaaaaeaii ia ?en.7. Ae?enii, o oeueiio aeiaaeeo niinoa??aa?oueny
aeaie?aoia i?enei?aiiy ?aaeoe??. Oe?eaai, ui oi?ia e?iaoe/ii? e?eai? –
aeaioaciaa. Oaeaee?noue neioaco aae?e-o?IE ciaioo?oueny c 2.5 iiieue/oa
a ia?o?e oac? aei 1.8 iiieue/oa o ae?oa?e. I?e a?aenooiino? iaaeeeoeo
EF-1 oaeaee?noue neioaco aae?e-o?IE noaiiaeea 1.25 iiieue/oa.

I?e cai?i? GTP ia GDP noeioeyoe?? aae?e-o?IE neioaoacii? aeoeaiino?
oaeoi?ii ia niinoa??aaeiny (?en.8), ui i?aeoaa?aeaeo? iiaeeea?noue
o/ano? eeane/iiai iio??eiiai eiiieaeno EF-1*GTP*ai?iiaoeee-o?IE a
iaoai?ci? noeioeyoe??. Ae?y GMPPNP — aiaeiaa GTP, ui ia
a?ae?ie?co?oueny, aoea aiaeia?/iith, oi/ ? iaioith ii aiie?ooae?, i?ae
ae?y GTP. Ciaioaiiy noeioeyoe?? aae?e-o?IE neioaoace a i?enooiino?
aiaeiaa GTP iiaeia iiynieoe cieaeaiiyi caeaoiino? noaiaeeieoeue EF-1H
eaoae?coaaoe oi?ioaaiiy EF-1-GTP aiane?aeie ooai?aiiy a?aeiinii
noaa?eueiiai eiiieaeno EF-1*GMPPNP, yeee aeaiaeeoue /anoeio iieaeoe
EF-1H

?en.7 Oaeaee?noue neioaco 14N-aae?e-o?IE aae?e-o?IE neioaoacith a
eiiieaen? c EF-1H a i?enooiino? (1) /e aac (2) EF-1. Eiioeaio?aoe?y
oa?iaioa a i?ia? — 0.6 iI. 50 iiieue EF-1 i?noeeiny a 100 iee
?ieoaaoe?eii? noi?o?.

?en.8 Aeoeai?noue aae?e-o?IE neioaoace a eiiieaen? c EF-1H a
i?enooiino? EF-1 (50 iiieue/100 iee noi?o?) oa GTP (1), GMPPNP (2) aai
GDP (3). (4) — aeoeai?noue oa?iaioa a i?enooiino? GMPPNP aac EF-1.
Eiioeaio?aoe?y aae?e-o?IE neioaoace 2.1 iI. Eiioeaio?aoe?? ioeeaioeae?a:
100 ieI GTP, 140 ieI GMPPNP, 100 ieI GDP.

c ?aaeoe?? (Janssen et al, 1988). Oe? aeai? aoee i?aeoaa?aeaeai? a iaoeo
aenia?eiaioao.

Niaoeeo?/i?noue aoaeoo EF-1 aoea ia?aa??aia cai?iith o aeuaiienai?e
nenoai? aoea??ioe/iiai oaeoi?a EF-1 ia i?iea??ioe/iee EF-Tu. Ine?eueee,
ye aeyaeeiny, EF-1 ia i?enei?th? iai?i ioeeaioeae?a a iieaeoe? EF-Tu,
niaoe?aeueio oaaao aoei i?eae?eaii ia?aaaaeaiith EF-Tu a GTP-ca’ycaio
oi?io, aeei?enoiaoth/e oinoiaiiei??oaao oa eiai e?iaco. Aeyaeeiny, ui
EF-Tu-GTP ia aieeaa? ia aeoeai?noue aae?e-o?IE neioaoace iaa?oue o
eiioeaio?aoe??, ui aoea a /ioe?e ?ace a?eueoa ca eiioeaio?aoe?th EF-1 a
aiaeia?/iiio aenia?eiaio? (?en.9). Ioaea, i?inoi nii??aeiaiino? aei
ai?iiaoeee-o?IE aeyaeeiny iaaeinoaoiuei aeey aeoeaaoe?? aae?e-o?IE
neioaoace.

Iaeiei ?c oaeoi?ia, ui ia? aieea ia noeioeyoi?iee aoaeo EF-1, aeyaeany
nooi?iue caaaa/aiiy noia?iiai i?aia?aoo o?IE aae?iiaith o?IE. I?e
ii??aiyii? i?aia?ao?a o?IE, ye? i?noeee 2.5% aai 20,4% o?IEVal,
aeyaeeiny, ui aeoeaaoe?y aae?e-o?IE neioaoace oaeoi?ii aeiiaaoe??
niinoa??aa?oueny o?eueee i?e aeei?enoaii? caaaa/aiiai ii o?IEVal
i?aia?aoo noia?ii? o?IE (?en.10). Ie iiynith?ii oea yaeua 10-e?aoiei
ciaioaiiyi (c 22000 aei 2500 iiieue ia oi/eo i?yii?) e?eueeino?
aeaaoeeeueiaaii? o?IE a caaaa/aiiio i?aia?ao?. sseui EF-1 caeaoiee
oi?ioaaoe iaeaiii?/iee eiieaen c aeaaoeeeueiaaiith o?IE (i?i ui aoaea
neacaii ieae/a), ?? cia/iee iaaeeeoie a ii??aiyii? c ai?iiaoeee-o?IE a
nenoai? i?ecaiaeeoue aei oiai, ui iaeaea an? iieaeoee oaeoi?a
ia?aaoaathoue a eiiieaen? c aeaaoeeeueiaaiith o?IE, oiaoi ia caeaoi?
aieeaaoe ia aeoeai?noue aae?e-o?IE neioaoace ca

?en.9 Oaeaee?noue neioaco aae?e-o?IE aae?e-o?IE neioaoacith a eiiieaen?
c EF-1H a i?enooiino? aaeoa??aeueiiai oaeoi?a aeiiaaoe?? EF-Tu. () —
aeiaeaii 50 iiieue EF-Tu/100 iee, () — aeiaeaii 200 iiieue EF-Tu/100
iee, () — aac EF-Tu. Eiioeaio?aoe?y aae?e-o?IE neioaoace — 1.7 iI.

?en. 10 Aeoeai?noue aae?e-o?IE neioaoace a i?enooiino? EF-1 (1,3) oa aac
iueiai (2,4) i?e aeei?enoaii? ye noano?ao?a i?aia?ao?a o?IE c ??ciei
ai?noii o?IEVal. A iaeiiio i?aia?ao? o?IE i?noeeiny 20,4% o?IEVal (1,2),
a ?ioiio — 2.5% o?IEVal (3,4). Aeey caaaa/aiiai i?aia?aoo e?euee?noue
o?IEVal ia oi/eo i?yii? aoea 13.1 iea, aeey noia?iiai — 13.8 iea.

?aooiie oi?ioaaiiy eeane/iiai iio??eiiai eiiieaeno c ai?iiaoeee-o?IE.

?ioei eiio?ieai aoei aeei?enoaiiy aae?e-o?IE neioaoace ?c ae??aeaeae?a.
Aeey oeueiai oa?iaioo ?ai?oa aoei anoaiiaeaii, ui noaae??th, yea e?i?oo?
oaeaee?noue, ? aeenioe?aoe?y neioaciaaii? ai?iiaoeee-o?IE. (Kern et al.,
1981). Aea aeiaeaaaiiy EF-1, iacaaaeath/e ia oa, ui oeae a?eie ia?
aenieo nii??aeiai?noue aei ai?iiaoeee-o?IE, ia aieeaaei ia neioac
aae?e-o?IE (?en.11) iaa?oue o i?enooiino? noaiaeeieoeue EF-1H, ye?
eaoae?cothoue GDP/GTP iai?i a iieaeoe? EF-1. Oaeei /eiii, iiaeia c?iaeoe
aeniiaie, ui i?enei?aiiy neioaco aae?e-o?IE aae?e-o?IE neioaoacith, yea
ciaoiaeeoueny a eiiieaen? c EF-1H, a i?enooiino? iaaeeeoeo

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0000000000000000000

?en.11. Aeoeai?noue aae?e-o?IE neioaoace c ae??aeaeae?a a i?enooiino?
(1) oa aac (2) EF-1. Eiioeaio?aoe?y aae?e-o?IE neioaoace — 1.6 iI.
E?euee?noue EF-1 50 iiieue/100iee noi?o?. E?ioeaaa eiioeaio?aoe?y EF-1 —
31 iI.

EF-1 ? niaoeeo?/iei aeey a?ee?a aeueo aoea??io ? iiaea aooe ?acoeueoaoii
a?eie-a?eeiaeo aca?iiae?e a oeueiio eiiieaen?.

sse? eiiiiiaioe eiiieaeno aae?e-o?IE neioaoaca — EF-1H aaaeeea? aeey
caaacia/aiiy ooieoe?iiaeueii? aca?iiae?? EF-1 c aae?e-o?IE neioaoacith?
Aeey a?aeiia?ae? ia oea ieoaiiy aoei aeioe?eueii i?iaanoe aeenioe?aoe?th
eiiieaeno ? iio?i eiai ?aeiino?oeoe?th a aeine?aeao in vitro c ie?aieo
eiiiiiaio?a, aea/ath/e aieea eiaeiiai c ieo ia noeioethth/ee aoaeo EF-1.
?ai?o aoei iieacaii, ui aae?e-o?IE neioaoaca,/anoeia N-e?ioeaaiai
aeiiaia yei? aeaa?aaeiaaia aeanoacith, ao?a/a? nii??aeiai?noue aei
EF-1H, aea caeeoa?oueny oaeith ae aeoeaiith a ai?iiaoeeethaaii? (Bec et
al, 1994). Ie i?iaeaiiino?oaaee, ui EF-1 i?ye ia aieeaa? ia aeoeai?noue
nei?i/aii? c N-e?ioey aae?e-o?IE neioaoace (?en.12a). Aea ia iiaeia
aeeeth/eoe, ui N-e?ioeaaa /anoeia iieaeoee oa?iaioo i?eeia? o/anoue o
aca?iiae?? c EF-1, oiaoi ?? aeaeaeaiiy iiaea i?ecaiaeeoe aei ao?aoe
oaei? aca?iiae??, ? ioaea aei ao?aoe noeioethth/iai aieeao EF-1 ia
aeoeai?noue aae?e-o?IE neioaoace. Oiio i?ney aeenioe?aoe?? eiiieaeno
aoei io?eiaii iaoeaio aae?e-o?IE neioaoaco ? aea/aii aoaeo EF-1 ia ??
aeoeai?noue. C ?en. 12a aeaeii, ui EF-1 ia aieeaa? ? ia aeoeai?noue
aeenioe?eiaaiiai c eiiieaeno iaoeaiiai oa?iaioo iaa?oue ca i?enooiino?
EF-1. Oaeei /eiii, anioe?aoe?y aae?e-o?IE neioaoace c EF-1H aaniethoii
iaiao?aeia aeey ooieoe?iiaeueii? aca?iiae?? oa?iaioo c EF-1. Ieoaiiy i?i
oa, yeee c eiiiiiaio?a EF-1H ? aaaeeeaei aeey oe??? aca?iiae??
caeeoa?oueny a?aee?eoei, oiio ui i? EF-1, i? EF-1, i? ?o eiia?iaoe?y ia
iaee i?yeiai aieeao ia aeoeai?noue ?iaeea?aeoaeueii? aae?e-o?IE
neioaoace ye a i?enooiino?, oae ? aac EF-1. Ia ci?ithaaea caaaeueii?
ea?oeie ? ci?ia ii?yaeeo aeiaaaeaiiy ??cieo noaiaeeieoeue EF-1H i?e
?aeiino?oeoe?? eiiieaeno. Iaiao?aeii caoaaaeeoe, ui ca?aeii c
e?oa?aoo?ieie aeaieie EF-1 ia? aeoaea aaeeeo caeaoi?noue aei
naiiaa?aaaoe?? oa ianiaoeeo?/ii? aa?aaaoe?? c ?ioeie eiiiiiaioaie
eiiieaeno (Bec et al, 1994), ? oiio ei?aeoia ?aeiino?oeoe?y eiiieaeno
EF-1H c ie?aieo eiiiiiaio?a iiaea aooe

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000000

?en.12 Aeoeai?noue ?iaeea?aeoaeueii? aae?e-o?IE neioaoace aac
N-e?ioeaai? /anoeie iieaeoee (A) oa ?iaeea?aeoaeueii? ?ioaeoii?
aae?e-o?IE neioaoace (A) a i?enooiino? (1) oa aac (2) EF-1
Eiioeaio?aoe?y iiaeeo?eiaaii? aae?e-o?IE neioaoace – 2.5 iI, ?ioaeoii? —
2.7 iI. 100 ieI GTP ? 110 iI EF-1 aoee i?enooi? a eiaei?e noi?o?. 50
iiieue EF-1 aoei aeiaeaii ia 100 iee ?ieoaaoe?eii? noi?o?.

oo?oaeiaia, yeui ia iaiiaeeeaa. Ine?eueee iaie aoei aeyaeaii, ui EF-1
cia/ii /ooeea?oee aei ae?? aeanoace, i?ae ?io? eiiiiiaioe eiiieaeno,
aeueoa?iaoeaiei oeyoii aecia/aiiy iiaeeeai? ?ie? EF-1 i?a ae aooe
/anoeiaee a?ae?ie?c aeanoacith eiiieaeno aae?e-o?IE neioaoaca-EF-1H. Aea
? oeae iaoiae ia aeaa /?oei? a?aeiia?ae?. Ioaea, caea?oueny eiia??iei,
ui aeey i?aeo?eiaiiy ooieoe?iiaeueii? aca?iiae?? aae?e-o?IE neioaoace c
EF-1H ? iaiao?aeiei eiiieaen aae?e-o?IE neioaoaca-EF-1H ye ?aeeia oe?ea.
?ieue ie?aieo eiiiiiaio?a EF-1H a caaacia/aii? aaciina?aaeiuei?
ia?aaea/? caiiai neioaciaaii? aae?e-o?IE a?ae aae?e-o?IE neioaoace aei
EF-1 iio?aao? iiaeaeueoiai aeine?aeaeaiiy.

Oaeei /eiii iaie anoaiiaeaii oaiiiai ooieoe?iiaeueii? aca?iiae?? oaeoi?a
aeiiaaoe?? o?aineyoe?? EF-1 c aae?e-o?IE neioaoacith. Noeioethaaiiy
aeoeaiino? neioaoace oaeoi?ii, i/aaeaeii, a?aeaoaa?oueny oeyoii
i?enei?aiiy aeenioe?aoe?? neioaciaaii? aae?e-o?IE c oa?iaioo ca ?aooiie
ia?aaea/? oe??? ai?iiaoeee-o?IE o iio??eiee eiiieaen
EF-1-GTP-aae?e-o?IE. Aeey cae?eniaiiy oaei? aca?iiae?? iaia’yceiaei ?
eiiieaeniooai?aiiy i?ae aae?e-o?IE neioaoacith oa EF-1H. ?acii c oei
iaie aoei iieacaii, ui a ee?oeiao nnaaoe?a /aiae?ia a?aeaoaa?oueny ca
o/ano? a?eueoino?, yeui ia an?o ai?iiaoeee-o?IE. Aeieea? ieoaiiy, /e
iiaeia aeyaeoe a aeine?aeao in vitro aca?iiae?th EF-1 ?c oeie
ai?iiaoeee-o?IE neioaoacaie, ye? ia aoiaeyoue aei neeaaeo ??cieo
aenieiiieaeoey?ieo eiiieaen?a? C?iaeoe oea aoei ae??oaii ia i?eeeaae?
oai?eaeai?e-o?IE neioaoace c ia/?iee e?iey.

Anoaiiaeaii, ui aeiaeaaaiiy EF-1-GDP aei ?ieoaaoe?eii? noi?o?
i?ecaiaeeei aei aeaie?aoiiai i?aeaeuaiiy aeoeaiino? oai?eaeai?e-o?IE
neioaoace (?en.13). Eaoae?oe/ia

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000

?en.13. Aeoeai?noue oai?eaeai?e-o?IE neioaoace c ia/?iee e?iey a
i?enooiino? (1) oa aac (2) 50 iiieue EF-1.. Ia’?i ?ieoaaoe?eii? noi?o? –
50 iee.

eiinoaioa (kcat) oa?iaioo a i?enooiino? EF-1 (0.76 nae-1) aoea a aeaa
?ace aeuith, i?ae aac iueiai (0.38 nae-1). Ne?ae iniaeeai a?aecia/eoe,
ui ia a?aei?io a?ae aae?e-o?IE neioaoace,

aeoeaaoe?y oai?eaeai?e-o?IE neioaoace a?aeaoaaeany i?ae ae??th GDP-oi?ie
EF-1. Oea aeicaieeei i?eionoeoe, ui iaoai?cie noeioethaaiiy oeeo aeaio
oa?iaio?a ??ci?, ? aeoeaaoe?y oai?eaeai?e-o?IE neioaoace iiaea aooe ia
ca’ycaia c oi?ioaaiiyi eeane/iiai iio??eiiai eiiieaeno
ai?iiaoeee-o?IE-EF-1-GTP, oiaoi c i?enei?aiiyi aeenioe?aoe??
ai?iiaoeee-o?IE c oa?iaioo, a a?aeaoaaoeny ca ?aooiie oaeoi?-neioaoacii?
aca?iiae?? ia aoaiao aei aeenioe?aoe?? neioaciaaii? ai?iiaoeee-o?IE.
Ae?enii, caeaaei?noue ??aiy aeoeaaoe?? a?ae eiioeaio?aoe?? EF-1 aoea
iiae?aia aei neaii?aeaeueii?, ui ? oeiiaei aeey aeinoa?e/ieo aca?iiae?e
(?en.14). Oea iiaea

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0000000000000000000

?en.14 Aeoeai?noue oaie?aeai?e-o?IE neioaoace a i?enooiino? ??cii?
e?eueeino? EF-1.. Ca 100% i?eeiyoi e?euee?noue oai?eaeai?e-o?IE (12
iiieue), neioaciaaii? aac o/ano? EF-1. Ia’?i ?ieoaaoe?eii? noi?o? – 50
iee.*an ?ieoaaoe?? – 2,5 oa.

na?ae/eoe ia ei?enoue aaciina?aaeiuei? aca?iiae?? oai?eaeai?e-o?IE c
EF-1. I?i niaoeeo?/i?noue aoaeoo aeoeaaoe?? na?ae/eoue iaci?ii?noue
oa?iaioaoeaii? aeoeaiino? a i?enooiino? ne?iaaoeiaiai aeueaoi?io aeea oa
i?iea??ioe/iiai oaeoi?a aeiiaaoe?? o?aineyoe?? EF-Tu. Aeey ia?aa??ee
i?eiouaiiy i?i oa, ui oaeoi? EF-1 aieeaa? ia ia?oo noaae?th ?aaeoe??
ai?iiaoeeethaaiiy, aoei aea/aii eiai ae?th ia AO?-i??ioinoaoiee iai?i,
yeee eaoae?co? oai?eaeai?e-o?IE neioaoaca. Anoaiiaeaii, ui oaeaee?noue
oe??? ?aaeoe?? oaeiae noeioeth?oueny oaeoi?ii aeiiaaoe?? EF-1 (?en.15).
Niinoa??aaeany oaeiae S-ia?aci?noue e?eai? caeaaeiino? aeoeaiino?
neioaoace a ?aaeoe?? AO?/??? iai?io a?ae eiioeaio?aoe?? EF-1.

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3005208b80482086504b208b804040000002d010500040000002d01010004000000f0010
600040000000201020005000000090200000000040000002d01040004000000f00103000
400000004010d000400000004010d0015000000fb024d000000000000009001000000000
3020112556b7261696e69616e426f646f6e69000079040000002d010300050000000102f
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00009000000fa0200000b000000000000002200040000002d010600070000001b04f1076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4000000f0010300040000000201020009000000fa0200000100000000000000220004000
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5000000090200000000040000002e010000040000002e011800030000001e0017000000f
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30017000000fb029a000000000000009001000000cc0302011254696d6573204e6577205
26f6d616e2043797200cc23040000002d010300050000000102ffffff000400000002010
10005000000090200000000040000002e010000040000002e011800030000001e0017000
000fb029a000000000000009001000000cc0302011254696d6573204e657720526f6d616
e2043797200cc23040000002d0106000500000009020000000012000000320a3309b8050
7000000d7e0f12c20f5e20058003c003c002200220044004000040000002d01040004000
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0000000000000000000

?en.15 Oaeaee?noue ATP-i??ioinoaoiiai iai?io, ui eaoae?co?
oai?eaeai?e-o?IE neioaoaca, a i?enooiino? (1) oa aac (2) 50 iiieue
EF-1. Ia’?i ?ieoaaoe?eii? noi?o? — 100 iee.

Oaeei /eiii, iiaeia c?iaeoe aeniiaie, ui noeioeyoi?iee aoaeo EF-1 ia
iia’ycaiee c i?enei?aiiyi aeenioe?aoe?? ai?iiaoeee-o?IE, ye o aeiaaeeo
aae?e-o?IE neioaoace. Oe?eeii eiia??ii, ui aca?iiae?y oaeoi?-neioaoaca
aai oaeoi? — o?IE — neioaoaca aaaea aei ci?ie eiioi?iaoe??
oai?eaeai?e-o?IE neioaoace, yea noa? a?eueo ni?eyoeeaith aei neioaco
oai?eaeai?eaaeai?eaoa, i?enei?thth/e oaeei /eiii ? caaaeueio oaeaee?noue
ai?iiaoeeethaaiiy. Aiai?y/e i?i iiaeeea? iaoai?cie noeioeyoe?? ne?ae
aeacaoe, ui aeey oai?eaeai?e-o?IE neioaoac ?c ae??aeaeae?a oa E.coli
oaeaee?noue-e?i?ooth/eie aoaiaie ? ye?nue i?ioeane, ui a?aeaoaathoueny
aei aeenioe?aoe?? ai?iiaoeee-o?IE c oa?iaioo (Fasiolo et al, 1978,
Baltzinger et al, 1982, Dibbelt et al, 1981). Oea na?ae/eoue ia ei?enoue
iaoiai aeniiaeo i?i i?e?iaeo noeioeyoe?? oaeoi?ii oai?eaeai?e-o?IE
neioaoace. Ine?eueee oeae aicei ia? /ioe?e noaiaeeieoe? oa aeaa aeoeaieo
oeaio?e, oa aa?o/e aei oaaae, ui a e?oa?aoo?? aaea aoei aenoiooi
i?eiouaiiy uiaei iiaeeeaino? eiiia?aoeaii? aca?iiae?? i?ae eaoae?oe/ieie
oeaio?aie noaiaeeieoeue (Dibbelt et al, 1981), ia ? ianiiae?aaiei
neaii?aeaeueiee oa?aeoa? e?eai? aieeao eiioeaio?aoe?? EF-1 ia neioac
oai?eaeai?e-o?IE ? ia ATP-i??ioinoaoiee iai?i, ui eaoae?co?
oai?eaeai?e-o?IE neioaoaca.

Oaeei /eiii, EF-1-GDP iiaea aca?iiae?yoe c oai?eaeai?e-o?IE neioaoacith
ua aei ai?iiaoeeethaaiiy o?IE oa i?ae /an oeueiai i?ioeano. ssea
ooieoe?iiaeueia cia/aiiy oe??? aca?iiae??? A iaaaaeai?e ?ai?oa noai?
/aiae?iaa oaeoi?a aeiiaaoe?? 1 a GTP-oi?i? a?aeaiaeeeany ?ieue
aeoeaioi?a ai?iiaoeee-o?IE i?ney ?? neioaco ai?iiaoeee-o?IE neioaoacith
(aeea. ?en.5). Nooo?a? aeieace ia ei?enoue oeueiai iaoai?cio io?eiai?
iaie ia i?eeeaae? aae?e-o?IE neioaoace. Noeioeyoe?y oai?eaeai?e-o?IE
neioaoace, i/aaeaeii, ia ? iane?aeeii ooieoe?iioaaiiy oeueiai iaoai?cio,
ai a?aeaoaa?oueny ia noaae?yo aei aeenioe?aoe?? ai?iiaoeee-o?IE c
oa?iaioo. Iaeiei c iaeiaio c’yniaaieo iiiaio?a o noai? /aiae?iaa o?IE a
a?eeiaiio neioac? ? oeyo i?ae A naeoii ?eainii oa ai?iiaoeee-o?IE
neioaoacaie. Iauiaeaaii aoei io?eiaii aeai?, i?i oa, ui aeaaoeeeueiaaia
o?IE, oae naii ye ? ai?iiaoeee-o?IE, ia ? a?eueiith o ee?oei?, a
caaaeaee ciaoiaeeoueny a eiiieaen? c ee?oeiieie no?oeoo?aie (Stapulionis
et al, 1995). ?icaeaath/e a?iioaco /aiae?iaa, ie cai?iiiioaaee, ui EF-1,
yeee ciaoiaeeoueny a GDP-oi?i? i?ney a?ae?ie?co GTP i?ae /an aca?iiae??
ai?iiaoeee-o?IE c A naeoii ?eainiie, caeaoiee ca’ycoaaoe aeaaoeeeueiaaio
o?IE a A naeo? ?eainiie, aeenioe?thaaoe c ?eainiie o neeaae? iio??eiiai
eiiieaeno EF-1-GDP-o?IE oa ia?aiineoe o?IE aei ai?iiaoeee-o?IE
neioaoace. A oeueiio aeiaaeeo aeyaeaia noeioeyoe?y GDP-oi?iith EF-1
ia?oi? noaae?? ?aaeoe?? ai?iiaoeeethaaiiy o?IE iiaea aooe i?inoei
a?aeca?eaeaiiyi aca?iiae?? iaeaiii?/iiai iio??eiiai eiiieaeno c
ai?iiaoeee-o?IE neioaoacith. Iaaia ??/, ui oea ia aeeeth/a? iiaeeeaino?
aca?iiae?? oaeoi?a ?c neioaoacith ? i?ney neioaco ai?iiaoeee-o?IE.

Aeey aeieaco oe??? aeiioace a ia?oo /a?ao iaiao?aeii iieacaoe ?nioaaiiy
iaeaiii?/iiai iio??eiiai eiiieaeno EF-1*GDP*o?IE. ?yae iai?yieo
niinoa?aaeaiue, ui aeacoaaee ia aeineoue aaeeeo eiia??i?noue oi?ioaaiiy
oaeiai eiiieaeno, aaea aoee c?iaeai? a oe?e ?iaio?. Aea iiaeeea?noue
ooai?aiiy eiiieaeno EF-1*GDP*o?IE aoea aia?oa i?yii aeieacaia iaie
iaoiaeii iaia?aiii? o?eueo?aoe??. Iaeii/anii aoei i?aeoaa?aeaeaii
iiaeeea?noue ooai?aiiy eiiieaeno EF-1*GTP*o?IE. Eiiieaen EF-1*GDP*o?IE
aoei ioa?aeoa?eciaaii oeoi?anoeaioiei aiae?cii. Aea/aeany
oeoi?anoeaioe?y i’yoe o?eioioai?eiaeo caeeoe?a a iieaeoe? EF-1, yea
iiaea ci?ithaaoeny i?e oi?ioaaii? eiiieaeno c o?IE. Ae?enii,
niinoa??aaeiny ca?eueoaiiy ?ioaineaiino? neaiaeo aaea a i?enooiino?
oaea?/? iaioi? e?eueeino? o?IE, i?ae e?euee?noue oaeoi?a (?en.16a).
Oe?eaai, ui i?aeaeuaiiy ?ioaineaiino? iiaeia aoei

?en. 16. Aieea o?IE ia ?ioaineai?noue ? iieiaeaiiy niaeo?a o?eioioaiiai?
oeoi?anoeaioe?? EF-1-GDP (A,C) oa o?eioioaiiaiai iio?aeiiai NaTa a
e?eueeino?, ui aoea aea?iiey?iith e?eueeino? o?eioioai?eiaeo caeeoe?a a
EF-1 (B, D). Aeey a?eueo /?oei? ?ethno?aoe?? niaeo?aeueiiai cnoao aeai?
(C, D) aoee ii?iae?ciaai? ii iaeneiaeuei?e ?ioaineaiino?.

?a?no?oaaoe iaa?oue ia oii? ianiaoeeo?/iiai ciaioaiiy ??aiy
oeoi?anoeaioe?? ca ?aooiie aaeni?aoe?? na?oea iieaeoeaie o?IE (?en.16b).
Oaeei /eiii, ?aaeueiee aieea o?IE ia aiaeiaaiio oeoi?anoeaioe?th aoa
aeaui a?eueoei, i?ae aeaoaeooaaeiny a aenia?eiaio?, i?iaaaeaiiio aac
o?aooaaiiy caeaoiino? o?IE iiaeeiaoe na?oei. C?inoaiiy ?ioaineaiino?
oeoi?anoeaioe?? a?aeia?aaea? ci?io ioi/aiiy oeoi?ioi??a a iieaeoe? EF-1,
oiaoi iiaea na?ae/eoe i?i eiioi?iaoe?eio ia?aaoaeiao a?eea i?e ooai?aii?
eiiieaeno c o?IE. A?eueo iaeiicia/iei na?ae/aiiyi oi?ioaaiiy oaeiai
eiiieaeno, ia ca’ycaiei c iiaeeeaeie a?oaoaeoaie ca ?aooiie
na?oeiiiaeeiaiiy o?IE, iiaea aooe cnoa iaeneioio niaeo?a aiaeiaaiii?
oeoi?anoeaioe?? a?eea i?e aeiaaaeaii? o?IE. Ae?enii, a eiio?ieueiiio
aenia?eiaio? ia niinoa??aaeiny i?yeiai aieeao o?IE ia iieiaeaiiy
iaeneioio oeoi?anoeaioe?? iiaeaeueiiai iaioeaeo NaTa, (?en. 16d), o oie
/an, eiee oa ae eiioeaio?aoe?y o?IE ni?e/eiyea iii?oiee (2ii) cnoa
iaeneioio oeoi?anoeaioe?? EF-1 o a?e ca?eueoaiiy aeiaaeeie oaee? (?en.
16c).

Ioaea, ci?ia iieiaeaiiy iaeneioio oa ?ioaineaiino? oeoi?anoeaioe?? EF-1
a i?enooiino? o?IE iaaeaicia/ii na?ae/eoue i?i ci?io eiioi?iaoe??
oaeoi?a aeiiaaoe?? a eiiieaen? c o?IE. A ui a?aeaoaa?oueny c iieaeoeith
o?IE a oaeiio eiiieaen?? A?aeiia?aeue ia oea ieoaiiy aoei io?eiaii ca
aeiiiiiaith iaoiae?a ?IEaciiai a?ae?ie?co oa o?i?/ii? iiaeeoeeaoe?? o?IE
??cii? niaoeeo?/iino?. Ea?oeia a?ae?ie?co o?IEPhe a i?enooiino? EF-1-GDP
ioeeaacith io?ooe eia?e iieacaia ia ?en. 17A. Aeyaeaii, ui EF-1-GDP
caoeua? a?ae ae?? ?IEace ioeeaioeaee a 68 oa 69 iieiaeaii? aeoeaioi?iiai
noaaea iieaeoee o?IE. E??i oiai, i?enooi?noue EF-1 i?ecaiaeeoue aei
iiyae iiaeo ?ic?ea?a a iiceoe?? 66 aeoeaioi?iiai noaaea ? a iic?oe?yo 73
oa 74 NNA-e?ioey iieaeoee. Oe?eaai, ui i?e cai?i? EF-1-GDP ia EF-1-GTP
ia a?aeaoaa?oueny ?iaeoeoe?? iiaeo ?ic?ea?a aai caoenoo ?ioeo
ioeeaioeae?a a iieaeoe? o?IE. Ia ?en.17A iieacaii ?acoeueoaoe a?ae?ie?co
o?IELeu ?IEacith O1. A i?enooiino? EF-1 ciinoa??aa?oueny i?aeneeaiiy
a?ae?ie?co iieaeoee o?IE i?ney G57 TC iaoe? oa G70 aeoeaioi?iiai noaaea,
a oaeiae aeayea i?aeneeaiiy ?ic?ea?a ia 3’ noi?ii? aa??aaaeueii? iaoe?.
Inoaii? iiaea aooe niaoe?oe/iei aeey aeiaaiiaoaeueieo o?IE, aei eeano
yeeo iaeaaeeoue o?IELeu. Ae?y ?IEace ?1, yea niaoeeo?/ia aei
iaeiini??aeueieo ae?eyiie iieaeoee o?IE, aoea iaeiaeiaith ye a
i?enooiino?, oae ? aac EF-1, oiaoi oe?eeii eiia??ii, ui a?eueo?noue
iaeiini??aeueieo /anoei iieaeoee o?IE ia aa?ooue o/ano? o aca?iiae?? c
EF-1.

Ia ?en.18 i?aaenoaaeaii ?acoeueoaoe o?i?/ii? iiae?o?eaoe?? o?IEPhe
aoeei?o?icina/iaeiith. EF-1 niaoeeo?/ii caoeua? a?ae aee?eoaaiiy
3’oinoaoi? a?oie ioeeaioeae?a G52, G53, A58, U68 oa C69, oiaoi /anoeie
aeoeaioi?iiai noaaea, TC-iaoe? oa TC–noaaea o?IEPhe. Ocaaaeueithth/e
aeai? ?IEaciiai a?ae?ie?co oa o?i?/ii? iiaeeo?eaoe??, iiaeia c?iaeoe
aeniiaie, ui oe? ?acoeueoaoe i?yii i?aeoaa?aeaeothoue ooai?aiiy
eiiieaeno EF-1-GDP-o?IE, oa aeicaieythoue ?aeaioeo?eoaaoe naeoe o?IE,
ye? i?eeiathoue o/anoue o aca?iiae?? c EF-1 (?en.19). ?iaeoeoe?y iiaeo
naeo?a a?ae?ie?co oa iineeaiiy aeayeeo ?nioth/eo ?ic?ea?a na?ae/eoue i?i
iaai? eiioi?iaoe?ei? ci?ie iieaeoee o?IE i?ae /an aca?iiae?? c EF-1.
Aa?oei

?en.17. A — aeaeo?ioi?aa?aia 32?-o?IEPhe, a?ae?ie?ciaaii? ioeeaacith
io?ooe eia?e (IIE) a i?enooiino? EF-1. 1 – i?ia?eaeeiiaee a?ae?ie?c, 2,6
— eiio?ie? aac ?IEace, ?ieoaaoe?y o?IE (2) aai o?IE+EF-1 (6), 3 –
iaiaaeaiee a?ae?ie?c T1 ?IEacith o?IEPhe, 4 – a?ae?ie?c IIE o?IEPhe, 5 –
a?ae?ie?c IIE o?IEPhe a eiiieaen? c EF-1-GDP, 7 – a?ae?ie?c IIE o?IEPhe
a eiiieaen? c EF-1-GOP, 8 – a?ae?ie?c IIE o?IEPhe a i?enooiino?
iaaeueaoi?ia. A — aeaeo?ioi?aa?aia 32?-o?IELeu, a?ae?ie?ciaaii? a
i?enooiino? EF-1-GDP ?eaiioeeaacith O1. 1 – a?ae?ie?c aeaii? o?IE a
i?enooiino? 7I na/iaeie i?e 550N, 2 – a?ae?ie?c o?IE aac aeiaaaie, 3, 5
– eiio?ieueia ?ieoaaoe?y o?IE (3) oa o?IE a i?enooiino? EF-1-GDP (5)
aac ?IEace, 4, 6 — a?ae?ie?c o?IE ?eaiioeeaacith O1 a i?enooiino?
EF-1-GDP (4) aai iaaeueaoi?ia (6) a?aeiia?aeii.

oaaae ? oa, ui ia?aaaaeaiiy iieaeoee EF-1 o GTP-oi?io, oi/ ?
noi?iaiaeaeoaaeiny aeayeith ci?iith aiie?ooaee i?ioaeoi?ii? ae?? oeueiai
a?eea ia a?ae?ie?c o?IE ioeeaacith io?ooe eia?e, aea ia aeeeeeaei iiyae
iiaeo naeo?a caoenoo ? iiaeo naeo?a a?ae?ie?co o?IE. Oea iiaea na?ae/eoe

?en.18 Ii??aiyiiy aeinooiiino? aoeei?o?icina/iaei? oinoaoieo a?oi
o?IEPhe a i?enooiino? EF-1-GDP oa aac iueiai. Aaee/eia R ?
ni?aa?aeiioaiiyi i?ae ?ioaineai?noth a?aeiia?aeieo aeaeo?ioi?aoe/ieo
nioa aee?eiaaii? o?IEPhe a i?enooiino? EF-1-GDP oa aac iueiai. Ci?ie.a
aeinooiiino? oinoaoieo a?oi o?IE aee?eoaaiith aaaaeaeeny ?noioieie, yeui
cia/aiiy R a?aeoeeyeiny a?ae 1 a?eueo i?ae ia 35%.

?en.19 Ae?eyiee o?IEPhe, ye? caoeuai? EF-1 a?ae ioeeaaciiai a?ae?ie?co
oa a?ae o?i?/ii? iiae?o?eaoe?? (aaeee? o?eeooieee), oa i?noey iineeaiiai
?IEaciiai a?ae?ie?co a i?enooiino? EF-1 (iae? o?eeooieee).

i?i iiae?aio eiioi?iaoe?th iieaeoee o?IE i?ae /an ?? aca?iiae?? c GDP-
aai c GOP- oi?iaie oaeoi?a.

Aeey c’ynoaaiiy i?inoi?iai? i?aai?caoe?? eiiieaeno EF-1-GDP-o?IE ie
aeei?enoaee aaea a?aeiio ca aeaieie ?aioaaiino?oeoo?iiai aiae?co
no?oeoo?o eeane/iiai iio??eiiai eiiieaeno i?iea??ioe/iiai oaeoi?a
aeiiaaoe?? EF-Tu, aiaeiaa GTP oa oai?eaeai?e-o?IE ae??aeaeae?a (Nissen
et al, 1995). I?e iaeeaaeaii? ia oeth no?oeoo?o aecia/aieo iaie naeo?a
aeaaoeeeueiaaii? o?IE, ui aa?ooue o/anoue o aca?iiae?? c oaeoi?ii, aoei
aeyaeaii i?aeoe/ii iiaiee ?o ca?a c ae?eyieaie iieaeoee ai?iiaoeee-o?IE,
ye? i?eeiathoue o/anoue o aca?iiae?? c i?iea??ioe/iei oaeoi?ii a
e?enoae? eiiieaeno (?en.20). Oe?eaai, ui oaeee ca?a niinoa??aa?oueny
iacaaaeath/e ia oa, ui ii??aiththoueny eiiieaen o?IE c GDP-oi?iith

?en.20. Noaiaoe/ia cia?aaeaiiy i?inoi?iai? no?oeoo?e iio??eiiai
eiiieaeno EF-1 *GDP * o?IE.

Ia a?aeii?e i?inoi?ia?e no?oeoo?? oai?eaeai?e-o?IE o neeaae? iio??eiiai
eiiieaeno c i?iea??ioe/iei oaeoi?ii EF-Tu oa GMPPNP iieacaii naeoe
aca?iiae?? aeaaoeeeueiaaii? o?IEPhe c aoea??ioe/iei oaeoi?ii EF-1.
E?oaaie iicia/ai? i?noey caoenoo o?IE, no??eeaie – i?noey iineeaiiy
a?ae?ie?co o?IE a i?enooiino? EF-1.

aoea??ioe/iiai oaeoi?a oa eiiieaen ai?iiaoeee-o?IE ?c GTP-oi?iith
i?iea??ioe/iiai oaeoi?a. Ioaea, ?nio? aeineoue aaeeea eiia??i?noue
?nioaaiiy a iieaeoe? EF-1 ni?eueiiai naeoa aeey ca’ycoaaiiy
ai?iiaoeee-o?IE ? o?IE, aai /anoeiaiai ia?ae?eooy oeeo naeo?a.

E?oa?aoo?i? aeai? na?ae/aoue i?i a?aenooi?noue cia/ii? eiioi?iaoe?eii?
??cieoe? i?ae GTP- oa GDP- oi?iaie EF-1 (van Damme et al, 1992). Ie, a
naith /a?ao, ia aeyaeee ?noioii? ??cieoe? i?ae ae?eyieaie aca?iiae??
o?IE c GTP- oa GDP-oi?iaie EF-1. A oaeiio ?ac? aecia/ai? iaie naeoe
aca?iiae?? o?IE c EF-1 iaiaaii aoaeooue a??ieie ? aeey eeane/iiai
iio??eiiai eiiieaeno EF-1*GTP*ai?iiaoeee-o?IE, oiaoi io?eiai?
?acoeueoaoe ? oaeoe/ii ia?oeie, ui aeathoue iaaio ?ioi?iaoe?th i?i
ae?eyiee o?IE, ye? i?eeiathoue o/anoue a oi?ioaaii? eeane/iiai
iio??eiiai eiiieaeno ai?iiaoeee-o?IE*EF-1*GTP. Ioaea, oe? ?acoeueoaoe ?
aeoaea aaaeeeaeie ? c oi/ee ci?o ?icoi?iiy iiae?iaeoeue caaaeueiiai
iaoai?cio aeiiaaoe?? iie?iaioeaeiiai eaioethaa o aeueo aoea??io.

Oaeei /eiii, i?aeoaa?aeaeaii ooai?aiiy iaeaiii?/iiai iio??eiiai
eiiieaeno EF-1*GTP*o?IE. Aea /e iiaea o?IE a oaeiio eiiieaen?
ia?aiineoeny aei ai?iiaoeee-o?IE neioaoace, ye cai?iiiiiaaii a iao?e
iiaeae? /aiae?iaa? Aeey aeieaco oeueiai iaiao?aeii aoei aeiaanoe
oi?ioaaiiy /aoaa?oeiiiai eiiieaeno EF-1*GDP*o?IE-ai?iiaoeee-o?IE
neioaoaca. Ii??aithth/e a?aeii? i?inoi?ia? no?oeoo?e eiiieaeno
oai?eaeai?e-o?IE neioaoace (Oh.termophilus) c o?IEPhe (Goldgur et al,
1997) oa eiiieaeno EF-Tu*GMPPNP*oai?eaeai?e-o?IE (Nissen et al, 1995),
iiaeia ia?ana?ae/eoeny o oiio, ui iaia? noa?e/ieo ia?aoeiae aeey
ooai?aiiy /aoaa?oeiiiai eiiieaeno. Aeey i?yii? aenia?eiaioaeueii?
ia?aa??ee iiaeeeaino? ?nioaaiiy oaeiai eiiieaeno aoei canoiniaaii iaoiae
“cao?eiee o aae?”, yeee aaco?oueny ia ci?i? ?ooeeaino? ioeea?iiai?
eeneioe i?ioyaii ?? aeaeo?ioi?aco a i?enooiino? a?ee?a, aei yeeo aiia
ia? nii??aeiai?noue (Sioud et al, 1996). Aoei aeei?enoaii aeaeo?ioi?ac a
aaa?ic?, yeee aeicaieya ?ooaoeny aeineoue aaeeeei iae?iiieaeoey?iei
eiiieaenai i?e iieacieeao ?I, ui iacia/ii a?ae??ciyeeny a?ae ?o
?ciaeaeo?e/ii? oi/ee. Aeaeo?ioi?aa?aia ia ?en. 21 aeicaiey? ioe?ieoe
ooai?aiiy ??cieo eiiieaen?a o?IEPhe. Aeaeii, ui niinoa??aa?oueny
cao?eiea ?ooo 32?-o?IEPhe ye iane?aeie ooai?aiiy eiiieaeno c EF-1-GDP
aai c oai?eaeai?e-o?IE neioaoacith.

?en. 21 Ooai?aiiy /aoaa?oeiiiai eiiieaeno
EF-1*GDP*o?IEPhe*oai?eaeai?e-o?IE neioaoaca. Iaaaaeaii ?acoeueoaoe
aeaeo?ioi?aco a 0.7% aaa?iciii aae?. 1 – o?IE, 2 – EF-1-GDP oa o?IE, 3 –
oai?eaeai?e-o?IE neioaoaca oa o?IE, 4 — EF-1-GDP, oai?eaeai?e-o?IE
neioaoaca oa o?IE.

Iaea?eueo aaaiiei ? oie oaeo, ui i?e ci?ooaaii? EF-1-GDP,
oai?eaeai?e-o?IE neioaoace oa o?IEPhe a?aeaoaa?oueny i/aaeaeiee cnoa
nioae o?IE a ua a?eueo “aenieiiieaeoey?io” ciio aaey, ui na?ae/eoue i?i
ooai?aiiy /aoaa?oeiiiai eiiieaeno EF-1*GDP*o?IEPhe*oai?eaeai?e-o?IE
neioaoaca. Oe?eaai, ui EF-1*GDP*o?IE ia ooai?th? /aoaa?oeiiee eiiieaen
i? c ae?oea?aeueaeaa?aeoinoaoaeaa?ae?iaaiacith, yea ia? oaeee aea ca?yae
(??~8.0), ai? c eaoaeacith, yea ia? oaeo ae iieaeoey?io iano (240
Eaea) ye oai?eaeai?e-o?IE neioaoaca. Oea na?ae/eoue i?i niaoeeo?/i?noue
ooai?aiiy eiiieaeno EF-1*GDP*o?IEPhe*oai?eaeai?e-o?IE neioaoaca. Ioaea,
i?aeoaa?aeaeaii iaoa i?eiouaiiy, ui i?e/eiith noeioeyoi?ii? ae?? EF-1 ia
aeoeai?noue oai?eaeai?e-o?IE neioaoace ? oi?ioaaiiy eiiieaeno oaeoi?a ?
oa?iaioo ua aei ii/aoeo ai?iiaoeeethaaiiy. Oaeee eiiieaen iiaea aieeaaoe
ia aca?iiae?th i?ae aeoeaieie oeaio?aie oai?eaeai?e-o?IE neioaoace,
i?enei?thth/e noaae?th neioaco oai?eaeai?eaaeai?eaoo, ? e??i oiai
caaacia/oaaoe iioeiaeueio i???ioaoe?th iieaeoee o?IE ii a?aeiioaiith aei
aeoeaiiai oeaio?a aiceio.

Eiioeaioe?y ooieoe?iiaeueii? eiiia?oiaioae?caoe?? a?eeiaiai neioaco o
aeueo aoea??io. Ocaaaeueithth/e ea?oeio, iiaeia c?iaeoe aeniiaie i?i
?nioaaiiy aeaio oeyo?a ia?aaea/? o?IE i?ae ai?iiaoeee-o?IE neioaoacith ?
EF-1. Aeaaoeeeueiaaia o?IE ia?aaea?oueny a?ae EF-1-GDP aei
ai?iiaoeee-o?IE neioaoace (iieacaii ia i?eeeaae? oai?eaeai?e-o?IE
neioaoace), a ai?iiaoeee-o?IE ia?aaea?oueny a?ae ai?iiaoeee-o?IE
neioaoace aei EF-1-GOP (iieacaii ia i?eeeaae? aae?e-o?IE neioaoace).
Aa?o/e oea aei oaaae, ie i?iiiio?ii oaeo iiaeaeue ooieoe?iioaaiiy
aia?aoo o?aineyoe?? a ?aaeei? /aiae?iaa o?IE (?en.22). EF-1 ? /iaieeii
i?ae ?eainiiaie ? a?iioaoe/iei iae?iiieaeoey?iei eiiieaenii, yeee
i?noeoue

?en. 22 A?iioaoe/ia iiaeaeue no?oeoo?ii-ooieoe?iiaeueii? i?aai?caoe??
o?aineyoe?eiiai eiiia?oiaioa

ai?iiaoeee-o?IE neioaoace oa EF-1H. Ca aeiiiiiaith EF-1 a?aeaoaa?oueny
aaciina?aaei? ia?aianaiiy ai?iiaoeee-o?IE a?ae oeueiai eiiieaeno aei
?eainiie oa ia?aianaiiy aeaaoeeeueiaaii? o?IE a?ae ?eainiie aei oeueiai
eiiieaeno. EF-1-GTP i?aoeth? a iai?yieo “ai?iiaoeee-o?IE neioaoaca —
?eainiia”, EF-1-GDP – a iai?yieo “?eainiia — ai?iiaoeee-o?IE neioaoaca”.
A oe?e iiaeae? /aiae?ia ai?iiaoeee-o?IE/o?IE caaacia/o?oueny
i?e?ieiiia?oiaioae?caoe??th EF-1, ai?iiaoeee-o?IE neioaoac, EF-1H,
?eainii oa i?IE a ee?oei?. Aeoaea oe?eaai, ui c iiaeae? aeieea?
iiaeeea?noue ni?aeiiia?oiaioae?caoe?? i?IE oa ioea o?IE c aioeeiaeiiaie,
ye? a?aeiia?aeathoue eiaeiiai i?IE ia oe?e ae?eyioe? (ia ?en.22
i?aaenoaaeaii no??eeaie), ui aeicaiey? cai?iiiioaaoe ye?nii iiaee
i?aeo?ae aei c’ynoaaiiy i?e/ei aeniei? oi/iino? o?aineyoe?? a
aoea??ioe/ieo ee?oeiao.

Ioaea, ?acoeueoaoe iaoeo aenia?eiaio?a na?ae/aoue i?i iayai?noue a
ee?oeiao nnaaoe?a, e??i no?oeoo?ii?, ua e ooieoe?iiaeueii?
eiiia?oiaioae?caoe?? aia?aoo a?eeiaiai neioaco. Ooieoe?iiaeueia
eiiia?oiaioae?caoe?y – oea oi?ioaaiiy o?aineyoe?eieo eiiia?oiaio?a, yea
iaiao?aeia aeey iioeiaeueiiai ooieoe?iioaaiiy aia?aoo a?eeiaiai neioaco
a ee?oeiao aeueo aoea??io. Aacoth/enue ia aeanieo oa e?oa?aoo?ieo aeaieo
ie i?iiiio?ii oaea aecia/aiiy o?aineyoe?eiiai eiiia?oiaioa.
O?aineyoe?eiee eiiia?oiaio – oea noeoii?noue (ainaiaeue)
iae?iiieaeoey?ieo eiiieaen?a, ye? ooai?ththoueny oa ?iciaaeathoueny i?ae
/an o?aineyoe?? i?IE ia ?eainiiao a iaiaaeaiiio ee?oeiiiio i?inoi??. Aei
neeaaeo eiiia?oiaioa aoiaeyoue o?IE, ai?iiaoeee-o?IE neioaoace, oaeoi?e
aeiiaaoe??, ?yae ?ioeo a?ee?a, oaeeo, iai?eeeaae, ye ianeioaoaci?
eiiiiiaioe aenieiiieaeoey?iiai eiiieaeno ai?iiaoeee-o?IE neioaoac, a
oaeiae i?IE oa ?eainiie. Ooieoe?iiaeueia cia/aiiy o?aineyoe?eiiai
eiiia?oiaioa aieiaiei /eiii iieyaa? a caaacia/aii? iiaeeeaino? i?yii?
ia?aaea/? noano?ao?a a?eeiaiai neioaco, cie?aia aeaaoeeeueiaaii? o?IE oa
ai?iiaoeee-o?IE, a?ae iaeiiai eiiieaeno aei ?ioiai a eaioethc?
iine?aeiaieo ?aaeoe?e i?ae /an o?aineyoe?? i?IE. No?oeoo?iith iniiaith
oi?ioaaiiy o?aineyoe?eieo eiiia?oiaio?a ie??i eiiiiiaio?a
a?eie-neioacoth/iai aia?aoo iiaeooue aooe aeoeiia? o?eaiaioe
oeeoineaeaoo, a oaeiae iaia?aie oi?noeiai aiaeiieaciaoe/iiai ?aoeeoeoio.

O ee?oei? o?aineyoe?ei? eiiia?oiaioe i?aai?ciaai? oaeei /eiii, ui
aeinooi yeciaaiieo o?IE, ye? ia ciaoiaeyoueny a oeae /an a eiiieaen? c
a?eeaie, ona?aaeeio eiiia?oiaioa ? oo?oaeiaiei. Oea a?aeaoaa?oueny
oiio, ui ai?iiaoeee-o?IE ? o?IE iino?eii ia?aaoaathoue o o?ce/iiio
ca’yceo c aoaeue-yeei ?c o?aineyoe?eieo eiiieaen?a. Aiaeii/an
oo?oaeiaiee ? aeo?ae iaciai? aiaeiaaieo o?IE, ye? aaea ooieoe?iiothoue a
eiiia?oiaio?. O?aineyoe?eiee eiiia?oiaio yaey? niaith aeineoue eaa?eueia
ooai?aiiy ? iiaea /anoeiai aai iiai?noth ?iciaaeaoeny i?e iiaiiio
caaa?oaii? o?aineyoe?? aeaii? i?IE /e ca iani?eyoeeaeo ciai?oi?o oiia,
ye? aeeeeeathoue coieieo a?eeiaiai neioaco.

Aeniiaee.

1. Iaaia oai?aoe/ia oyaeaiiy i?i iiaeeea?noue eiiia?oiaioae?caoe??
aia?aoo a?eeiaiai neioaco aeueo aoea??io ia aoei i?aeoaa?aeaeaii ia
ii/aoie iaoeo aeine?aeaeaiue i?yeith aenia?eiaioaeueiith ?ioi?iaoe??th
uiaei ooieoe?iiaeueiiai cia/aiiy oeueiai oaiiiaio. A i?aaenoaaeai?e
aeena?oaoe?? io?eiaii a?oioiai? aenia?eiaioaeuei? aeai?, ye? aeicaieeee
?ic?iaeo eiioeaioe?? ooieoe?iiaeueii? eiiia?oiaioae?caoe?? aia?aoo
o?aineyoe?? oa a?iioaoe/ii? iiaeae? no?oeoo?ii-ooieoe?iiaeueii?
i?aai?caoe?? o?aineyoe?eiiai eiiia?oiaioo. Aia?oa cai?iiiiiaaii noaio
/aiae?iaa o?IE/ai?iiaoeee-o?IE, yea aeaoaeueii iieno? ??ci? aa??aioe
ia?aaea/? o?IE i?ae /an neioaco iie?iaioeaeiiai eaioethaa. Iieacaii
iioaioe?eia ?aaoeyoi?ia cia/aiiy ooieoe?iiaeueii? eiiia?oiaioae?caoe??,
o oiio /ene? cai?iiiiiaaii ?? iiaeeeao ?ieue a caaacia/aii? oi/iino?
o?aineyoe?? a ee?oeiao aeueo aoea??io.

2. Aia?oa ?ic?iaeaii oa aeei?enoaii nenoaio ni?yaeaii?
ia?iaaa?e?caoe??-o?aineyoe?? a ee?oeiao oaa?ei oa ethaeeie, ui
caaacia/o? oaeo ae oaeaee?noue a?eeiaiai neioaco, ye ? ?ioaeoi? ee?oeie.
?ic?iaeaii iaoiaee aeae?eaiiy oaeoi?a aeiiaaoe?? 1 oa oai?eaeai?e-o?IE
neioaoace c ia/?iee e?iey.

3. Aeyaeaii no?oeoo?ii-ooieoe?iiaeueio eiiia?oiaioae?caoe?th
ai?iiaoeee-o?IE a ee?oeiao aeueo aoea??io:

a) a aenia?eiaioao c iiaea?eiith i?oeith iieacaii, ui aiaeiaaiia
ai?iiaoeee-o?IE ia aeoiaeeoue iaciai? c ia?iaaa?e?ciaaieo ee?oei
eeoaenueeiai oii’y/ea, oiae? ye ?iciiae?e aeciaaiii aeiaeaii?
ai?iiaoeee-o?IE i?ae ee?oeiiei oa iicaee?oeiiei i?inoi?aie ?
i?iii?oe?eiei ia’?iai oeeo i?inoi??a;

a) anoaiiaeaii, ui aiaeiaaiia ai?iiaoeee-o?IE, ia a?aei?io a?ae
aeciaaiii aeiaeaii?, caoeuaia a?ae a?ae?ie?co aeiaaaeaiith ciai?
?IEacith a ia?iaaa?e?ciaaieo ee?oeiao;

a) aeyaeaii, ui aeiaeaia ciai? ai?iiaoeee-o?IE ia iiaea aoaeoeaii
aeei?enoiaoaaoenue aia?aoii o?aineyoe?? ia?iaaa?e?ciaaieo ee?oei, a oie
/an, eiee aiaeiaaiia ai?iiaoeee-o?IE aa?a aeoeaio o/anoue a o?aineyoe??.

4. Ia i?eeeaae? ia?iaaa?e?ciaaieo o?a?iaeano?a ethaeeie iieacaii
iiaeeea?noue caeaaeiiai a?ae ai?noo iae?ia?a?a ?iciaaeo o?aineyoe?eieo
eiiia?oiaio?a:

a) iaoiaeii oeoi?anoeaioii? i?e?ineii?? i?iaeaiiino?iaaii ci?io
aioo??oiueiee?oeiii? eieae?caoe?? oaeoi?a aeiiaaoe?? EF-1 a caeaaeiino?
a?ae oinoie?aaoeio;

a) ia aeyaeaii aieeao oinoie?aaoeio ia eieae?caoe?th ?ioi? noaiaeeieoe?
oaeoi?a aeiiaaoe?? EF-1;

a) cai?iiiiiaaii ?nioaaiiy iiaiai ??aiy ?aaoeyoe?? a?eeiaiai neioaco –
oeyoii anioe?aoe??-aeenioe?aoe?? o?aineyoe?eieo eiiia?oiaio?a;

a) i?eiouaii, ui iaeiei ?c iina?aaeiee?a a oaeiio oei? ?aaoeyoe??? ?
iaaeoeai? eiioi?ia?e o?IE, ye? ooai?ththoueny a i?aai?ci? ca
aeno?aiaeueieo oiia oa iathoue ?ia?a?oi?iee aieea ia o?aineyoe?th i?IE.

5. Aecia/aii ooieoe?iiaeueio ?ieue noaa?eueiiai eiiieaeno aae?e-o?IE
neioaoaca-EF-1H a ee?oeiao aeueo aoea??io:

a) iieacaii, ui EF-1-GTP noeioeth? ii/aoeiao oaeaee?noue neioaco
aae?e-o?IE aae?e-o?IE neioaoacith, yea ia?aaoaa? a eiiieaen? c
ioeueoenoaiaeeie/iith oi?iith oaeoi?a aeiiaaoe?? EF-1 (EF-1H).
A?aenooi?noue aoaeoo GDP-oi?ie oaeoi?a na?ae/eoue i?i o/anoue a
iaoai?ci? noeioeyoe?? eeane/iiai iio??eiiai eiiieaeno
EF-1*GTP*ai?iiaoeee-o?IE;

a) i?iaeaiiino?iaaii niaoeeo?/i?noue ooieoe?iiaeueii? aca?iiae?? EF-1 c
aae?e-o?IE neioaoacith aeey a?ee?a aeueo aoea??io;

a) aeyaeaii aaniethoio iaiao?aei?noue ia?aaoaaiiy aae?e-o?IE neioaoace a
eiiieaen? c EF-1H aeey ?? ooieoe?iiaeueii? aca?iiae?? c EF-1, ui
na?ae/eoue i?i aecia/aeueio ?ieue oeueiai eiiieaeno a caaacia/aii?
/aiae?iaa aae?e-o?IE.

6. Cai?iiiiiaaii ? aenia?eiaioaeueii aeiaaaeaii iiaeeea?noue i?yii?
ia?aaea/? aeaaoeeeueiaaii? o?IE a?ae EF-1 aei ai?iiaoeee-o?IE neioaoace:

a) i?iaeaiiino?iaaii ?noioiee noeioeyoi?iee aoaeo EF-1-GDP ia
aeoeai?noue oai?eaeai?e-o?IE neioaoace a aeoeaaoe?? ai?iieeneioe oa
ai?iiaoeeethaaii? o?IE;

a) ??cieie iaoiaeaie aeieacaii ooai?aiiy iaeaiii?/iiai iio??eiiai
eiiieaeno EF-1*GDP*o?IE;

a) i?iaeaiiino?iaaii ooai?aiiy /aoaa?oeiiiai eiiieaeno
EF-1*GDP*o?IE*oai?eaeai?e-o?IE neioaoaca.

7. ?ic?iaeaii eiioeaioe?th ooieoe?iiaeueii? eiiia?oiaioae?caoe??
a?eie-neioacoth/iai aia?aoo aeueo aoea??io oa a?iioaoe/io iiaeaeue
no?oeoo?ii-ooieoe?iiaeueii? i?aai?caoe?? o?aineyoe?eiiai eiiia?oiaioo.
Cie?aia, aenoiooi aenia?eiaioaeueii iaa?oioiaaia iieiaeaiiy i?i
i?ioeeaaeii iai?aaeaio ae?th eiiieaen?a EF-1-GDP oa EF-1-GTP a
aeinoaaoe? o?IE a?ae ?eainiie aei ai?iiaoeee-o?IE neioaoace oa
aeinoaaoe? ai?iiaoeee-o?IE a?ae ai?iiaoeee-o?IE neioaoace aei ?eainiie
a?aeiia?aeii.

IA?AE?E ?IA?O, UI IIOAEEEIAAI? CA OAIITH AeENA?OAOe??.

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protein synthesis in vivo. // Proc. Natl. Acad. Sci. USA. — 1991. — 88.
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4. Negrutskii B.S., Budkevich T.V., Shalak V.F., Turkovskaya G.V.,
El’skaya A.V. Rabbit translation elongation factor 1 stimulates the
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5. Petrushenko Z.M., Negrutskii B.S., Ladokhin A.S., Budkevich T.V.,
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7. Negrutskii B.S., Shalak V.F., Kerjan P., El’skaya A.V., Mirande M.
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8. Iaa?ooeeee A.N. O?ainii?oiay ?IE eae oaeoi? ?aaoeyoeee aaeeiaiai
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10. Iaa?ooeeee A.N. Aeeyiea oinoie?aaoeia ia ?ani?aaeaeaiea ?acee/iuo
noauaaeeieoe oaeoi?a yeiiaaoeee o?aineyoeee 1 a ia?iaaaeeece?iaaiiuo
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inactive tRNA conformers and leucyl-tRNA synthetase from rabbit liver.
// Eur. J. Biochem.-1987.-164.- P. 56-59.

17. Iaa?ooeeee A.N., Aeueneay A.A. Acaeiiaeaenoaea ?acee/iuo eiioi?ia?ia
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ia/aie e?ieeea. // No?oeoo?a e aaiaoe/aneea ooieoeee
aeiiieeia?ia.-Eeaa.-Iaoe.aeoiea.-1981.-C. 36-38.

19. Iaa?ooeeee A.N. Nenoaia nii?yaeaiiie ia?iaaaeeecaoeee-o?aineyoeee a
eeaoeao aeeaioiuo e /aeiaaea // Iieaeoey?iay aaiaoeea e aeioaoiieiaey. —
Ieine.- 1998.-N. 236-238.

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aeiiieeia?ia”. — Eueaia. — 1988. — N.88.

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translation machinery // Proc. International Conf. “Frontiers in
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Conf. “Frontiers in translation”. — Victoria (Canada). — 1995. — P.169.

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Madison (USA). — 1995. — P. 185.

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17th Inter. tRNA Workshop. — Shiba (Japan). -1997. — P.8-26.

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different subunits of translation elongation factor 1 in gently
permeabilized human fibroblasts. // Abstr. of 17th Inter. tRNA Workshop.
— Shiba (Japan). -1997. — P.8-17.

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Channeling of aminoacyl-tRNA in the valyl-tRNA synthetase:EF-1H complex
// Abstr. of EMBO Workshop on Structure and Function of aminoacyl-tRNA
synthetases. — Mittelwihr(France). — 1998. — P.114.

27. El’skaya A.V., Petrushenko Z.M., Negrutskii B.S., Budkevich T.V.,
Shalak V.F. Eukaryotic aminoacyl-tRNA synthetases and EF-1 as possible
partners in tRNA channeling // Abstr. of EMBO Workshop on Structure and
Function of aminoacyl-tRNA synthetases. — Mittelwihr(France). — 1998. —
P.40.

Negrutskii B. S. Functional compartmentalization of the mammalian
translation machinery. – Manuscript. Thesis for Doctor of Biology
degree. Speciality 03.00.03 — Molecular biology. Institute of Molecular
Biology and Genetics, National Academy of Sciences of Ukraine, Kiev,
1999.

Functional compartmentalization of the protein synthetic machinery in
higher eukaryotes was studied using permeabilized with saponin or
electroporated cells. For the first time the system of coupled
permebilization-translation was developed for CHO cells and human
fibroblasts demonstrating the same efficiency of translation process in
intact and permeabilized cells. A possibility of compartmentalization of
aminoacyl-tRNA (aa-tRNA) in CHO cells was tested by studying aa-tRNA
distribution between intracellular and extracellular spaces of
permeabilized cells. In double-label experiments 25-fold retardation
inside permeabilized cells was observed for endogenous aa-tRNA while the
ratio of exogenous aa-tRNA present inside and outside was similar to
that of intracellular and extracellular volumes. Moreover, cellular
aa-tRNA was protected against action of added RNAse A while exogenous
aa-tRNA was easily hydrolysed by RNAse inside the cells. After
homogenization of the cells no protection of endogenous aa-tRNA against
RNAse was found. Thus, both distribution and RNAse protection
experiments show structural compartmentalization of cellular
aminoacyl-tRNA.

One of the important mechanisms to realize potential advantages of
compartmentalization may be channeling, or direct transfer of
intermediates from one component to another in a metabolic chain.
Besides the need for some structural organization, another evidence of
channeling mechanism is the impossibility of exogenous intermediates to
enter a channeled pathway. Thus, the level of utilization of cellular
and exogenously added aa-tRNA for protein synthesis in perforated cells
was studied. Contrary to a cell-free system, in which no advantage was
found, about 15-fold preference for cellular aa-tRNA was detected in
permeabilized cells, i.e. endogenous aa-tRNA was much more efficient
precursor, than exogenous aa-tRNA, for protein biosynthesis.

Thus, both structural and functional criteria for channeling are
fulfilled in eukaryotic protein synthesis. We suggested that channeling
of tRNA occurs in some labile structures – translational compartments
which are destroyed during homogenization. To obtain the information
about such compartments, namely, about possible changes in their
organization under different cell conditions, the fluorescently labeled
and subunits of elongation factor 1 (EF-1) were used as compartment
markers. Distribution of proteins in permeabilized human fibroblasts
VH25 was compared under regular and low energy (no phosphocreatine)
conditions. The absence of phosphocreatine in the incubation mixture was
shown to cause almost complete stop of translation. The same granular
EF-1 localization in cytoplasm was found under normal and low-energy
conditions while the cytoplasmic distribution of EF-1 was more granular
under normal conditions (resembling that of EF-1) and was much more
nucleus-concentrated under low energy conditions. Thus, it is shown that
involvement of the important component of translation apparatus — EF-1 —
into suggested translational compartments may depend on the efficacy of
protein biosynthesis in cell. Difference in EF-1 and EF-1 localization
under low energy conditions may reflect different levels of
immobilization of these proteins inside the cells, as earlier suggested.
As far as we know these data are the first to reveal the correlation
between depletion of energetic compound and the altered localization of
a translation machinery component in eukaryotic cells.

The possibility of aminoacyl-tRNA synthetase-EF-1 interaction, suggested
by channeling hypothesis, was studied in vitro using rabbit liver EF-1,
phenylalanyl-tRNA synthetase (PheRS) and valyl-tRNA synthetase
(ValRS)-EF-1H complex. EF-1 caused 2-fold stimulation of both valyl-tRNA
and phenylalanyl-tRNA formation. The stimulation was rather specific
since BSA and bacterial factor EF-Tu had no stimulatory capacity.
Nevertheless, the mechanisms of EF-1-induced stimulation were quite
different for two synthetases. Stimulation of ValRS required GTP
strongly suggesting classical ternary complex valyl-tRNA-GTP-EF-1
involvement. In that case EF-1 stimulatory effect appears to occur at
the stage of aminoacyl-tRNA dissociation from the synthetase as
suggested by channeling hypothesis. Interestingly, ValRS separated from
EF-1H cannot be activated by EF-1 demonstrating the absolute requirement
of the structure of whole valyl-tRNA synthetase-EF-1H complex for the
stimulation. On the contrary, activation of PheRS can be induced by GDP
form of EF-1 as well. Moreover, the stimulatory effect was observed
already at the level of phenylalanine activation, suggesting
factor-synthetase interaction to occur at some step before
aminoacyl-tRNA dissociation. A special functional role was suggested for
EF-1-GDP complex, namely, to form EF-1*GDP*tRNA complex and to deliver
deacylated tRNA to aminoacyl-tRNA synthetase.

The ability of EF-1-GDP to form a complex with tRNA was proved by
several independent techniques. Analysis of EF-1-GDP endogenous
fluorescence showed possible conformational change of the protein in the
presence of tRNAPhe. On the other side, partial RNAse hydrolysis and
chemical modification studies indicated conformational changes of
tRNAPhe and tRNALeu in the presence of EF-1-GDP. Interestingly, the
sites of tRNA interaction with EF-1-GDP coincided well with known
aminoacyl-tRNA sites of interaction with bacterial factor EF-Tu-GTP
suggesting similarity of both complexes. Finally, the formation of
quaternary complex EF-1-GDP-tRNAPhe-PheRS was demonstrated by “band
shift” technique. Thus, the principal possibility of two ways to
transport tRNA between aminoacyl-tRNA synthetase and EF-1 was
demonstrated. Deacylated tRNA can be delivered from EF-1-GDP to
synthetase (as in the case of PheRS), and aminoacyl-tRNA can be
delivered from synthetase to EF-1-GTP (as in the case of ValRS). On the
basis of these findings a model of translation apparatus functioning is
developed which suggests EF-1 to be a shuttle between ribosome and
hypotetic macromolecular complex of synthetases and EF-1H.

The concept of functional compartmentalization of mammalian translation
apparatus is put forward. Functional compartmentalization is the
formation of labile protein-protein and protein-RNA complexes which
provide organization of the translation compartments and their efficient
functioning. The definition for translational compartment is as follows:
it is an assembly of macromolecular complexes which are dynamically
converted during mRNA translation in some limited space.

Iaa?ooeeee A.N. Ooieoeeiiaeueiay eiiia?oiaioaeecaoeey aiia?aoa
o?aineyoeee o aunoeo yoea?eio. – ?oeiienue. Aeenna?oaoeey ia nieneaiea
o/aiie noaiaie aeieoi?a aeieiae/aneeo iaoe ii niaoeeaeueiinoe 03.00.03
iieaeoey?iay aeieiaey. EIAeA IAI Oe?aeiu, Eeaa, 1999 a.

Iieacaia no?oeoo?iay e ooieoeeiiaeueiay eiiia?oiaioaeecaoeey
aieiiaoeee-o?IE a eeaoeao ieaeiieoathueo, iaania/eaathuay
ooieoeeiie?iaaiea iaoaiecia /aiaeeiaa o?IE i?e o?aineyoeee. Iaia?oaeaia
caaeneiinoue aioo?eeeaoi/iie eieaeecaoeee oaeoi?a yeiiaaoeee EF-1, ii ia
EF-1, oo oinoie?aaoeia – iniiaiiai eiiiiiaioa AO?-?aaaia?e?othuae
nenoaiu. I?iaeaiiino?e?iaaii ooieoeeiiaeueiia acaeiiaeaenoaea EF-1-GDP n
oaieeaeaiee-o?IE neioaoacie, e EF-1-GTP – c aaeee-o?IE neioaoacie,
iaoiaeyuaeny a eiiieaena n EF-1H. Auaeaeiooi i?aaeiieiaeaiea i
?aciiiai?aaeaiiii aeaenoaee yoeo oi?i EF-1: EF-1-GDP aeinoaaeyao
aeaaoeeee?iaaiioth o?IE e aieiiaoeee-o?IE neioaoaca, a EF-1-GTP
aeinoaaeyao aieiiaoeee-o?IE io neioaoacu e ?eainiia. Iieacaii
oi?ie?iaaiea eiiieaena EF-1-GDP-o?IEPhe-oaieeaeaiee-o?IE neioaoaca. Ia
iniiaaiee iieo/aiiuo aeaiiuo ?ac?aaioaia eiioeaioeey ooieoeeiiaeueiie
eiiia?oiaioaeecaoeee aiia?aoa o?aineyoeee ieaeiieoathueo, auaeaeiooi
iieiaeaiea i o?aineyoeeiiiii eiiia?oiaioa.

Eeth/aaua neiaa: yoea?eioe/aneee aeineioac aaeea, /aiaeeia,
eiiia?oiaioaeecaoeey, aieiiaoeee-o?IE neioaoacu, oaeoi? yeiiaaoeee 1.

Iaa?ooeueeee A.N. Ooieoe?iiaeueia eiiia?oiaioae?caoe?y aia?aoo
o?aineyoe?? o aeueo aoea??io. – ?oeiien. Aeena?oaoe?y ia caeiaoooy
a/aiiai nooiaith aeieoi?a a?ieia?/ieo iaoe c? niaoe?aeueiino? 03.00.03
iieaeoey?ia a?ieia?y. ?IA?A IAI Oe?a?ie, Ee?a, 1999 ?.

Iieacaii no?oeoo?io oa ooieoe?iiaeueio eiiia?oiaioae?caoe?th
ai?iiaoeee-o?IE a ee?oeiao nnaaoe?a, ui caaacia/o? ooieoe?iioaaiiy
iaoai?cio /aiae?iaa o?IE i?ae /an o?aineyoe??. Ciaeaeaii caeaaei?noue
aioo?eoiueiee?oeiii? eieae?caoe?? oaeoi?a aeiiaaoe?? EF-1, aea ia EF-1,
a?ae oinoie?aaoeio – iniiaiiai eiiiiiaioa AO?-?aaaia?oth/i? nenoaie.
I?iaeaiiino?iaaii ooieoe?iiaeueio aca?iiae?th EF-1-GDP c
oai?eaeai?e-o?IE neioaoacith, oa EF-1-GTP – c aae?e-o?IE neioaoacith, ui
ciaoiaeeoueny a eiiieaen? c EF-1H. Aenoiooi i?eiouaiiy i?i
i?ioeeaaeiini?yiiaaio ae?th oeeo oi?i EF-1: EF-1-GDP aeinoaaey?
aeaaoeeeueiaaio o?IE aei ai?iiaoeee-o?IE neioaoace, a EF-1-GTP
aeinoaaey? ai?iiaoeee-o?IE a?ae neioaoace aei ?eainiie. Iieacaii
oi?ioaaiiy eiiieaena EF-1-GDP-o?IEPhe-oai?eaeai?e-o?IE neioaoaca.
Aacoth/enue ia io?eiaieo aeaieo, ?ic?iaeaii eiioeaioe?th
ooieoe?iiaeueii? eiiia?oiaioae?caoe?? aia?aoo o?aineyoe?? nnaaoe?a,
aenoiooi iieiaeaiiy i?i o?aineyoe?eiee eiiia?oiaio.

Eeth/ia? neiaa: aoea??ioe/iee a?ineioac a?eea, /aiae?ia,
eiiia?oiaioae?caoe?y, ai?iiaoeee-o?IE neioaoace, oaeoi? aeiiaaoe?? 1.

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