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

?INOEOOO IIEAEOEss?II? A?IEIA?? OA AAIAOEEE

Ia i?aaao ?oeiieno

NA?AA?ssIEE Ieaenaiae? ?ee?/

OAeE 577.217.34

577.217.344

577.5.53

577.217.39

?EAINIIIA AO?aca AEUEO AOEA??IO

03.00.03 — iieaeoey?ia a?ieia?y

Aaoi?aoa?ao

aeena?oaoe?? ia caeiaoooy iaoeiaiai nooiaiy

eaiaeeaeaoa a?ieia?/ieo iaoe

Ee?a — 1998

Aeena?oaoe??th ? ?oeiien

?iaioa aeeiiaia o a?aeae?e? iaoai?ci?a o?aineyoe?? aaiaoe/ii?
?ioi?iaoe?? ?inoeoooo iieaeoey?ii? a?ieia?? oa aaiaoeee IAI Oe?a?ie (i.
Ee?a).

Iaoeiaee ea??aiee:

aeieoi? a?ieia?/ieo iaoe, i?ioani?,

aeaaeai?e IAI Oe?a?ie

?EUeNUeEA Aaiia Aaeaioei?aia,

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

a?aeae?eii iaoai?ci?a o?aineyoe?? aaiaoe/ii? ?ioi?iaoe??

Io?oe?ei? iiiiaioe:

aeieoi? a?ieia?/ieo iaoe,

Eiceia Aaeoa?ae Aiae??eiae/

?inoeooo iieaeoey?ii? a?ieia?? oa aaiaoeee IAI Oe?a?ie,

i?ia?aeiee iaoeiaee ni?a?ia?oiee a?aeae?eo aiceiieia?? a?eeiaiai
neioaco

aeieoi? a?ieia?/ieo iaoe, i?ioani?

Noa?iaeoa Ieeiea Oaaei?iae/

?inoeooo a?io?i?? ?i. I.A. Iaeeaae?ia IAI Oe?a?ie, caa.

a?aeae?eii a?io?i?? naini?ieo ? ?aaoeyoi?ieo nenoai.

I?ia?aeia onoaiiaa:

Ee?anueeee oi?aa?neoao ?iai? Oa?ana Oaa/aiea, eaoaae?a a?io?i??

Caoeno aeena?oaoe?? a?aeaoaeaoueny 23 /a?aiy 1998 ?. i 10-00 aiae. ia
can?aeaii? niaoe?ae?ciaaii? a/aii? ?aaee Ae 26.237.01 a ?inoeooo?
iieaeoey?ii? a?ieia?? oa aaiaoeee IAI Oe?a?ie ca aae?anith: 252143, i.
Ee?a-143, aoe. Caaieioiiai, 150.

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

Aaoi?aoa?ao ?ic?neaii 23 o?aaiy 1998 ?.

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

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

CAAAEUeIA OA?AEOA?ENOEEA ?IAIOE

Aeooaeuei?noue i?iaeaie. A?aeiii, ui oeeee aeiiaaoe?? iie?iaioeaeiiai
eaioethao ia ?eainiiao a on?o aeeaeo i?aai?ciao i?iiioo?oueny aeaiia
a?eeiaeie oaeoi?aie. I?ioa aeyaeeiny, ui ??ci? aeaee a?ea?a i?noyoue ua
iaeei aeiaeaoeiaee ?ic/eiiee oaeoi? EF-3(Skogerson, 1979),
ooieoe?iioaaiiy yeiai ? oaeiae nooo?aei aeey noaae?? aeiiaaoe??. Ia
a?aei?io a?ae aeaio iiia?aaei?o oaeoi??a EF-3 ia? neaaeo AO?acio
aeoeai?noue, yea ia aeae?eueea ii?yaee?a ca?eueoo?oueny o i?enooiino?
?eainii. Aeaoaeueia aea/aiiy o/ano? EF-3 o a?ineioac? a?eea iaaeaei
i?aenoaao i?eionoeoe, ui a?i i?iiioo? ia?ao?ae ?eainiie c iino- o
i?ao?aineieaoe?eiee noai (Nierhaus, 1993) oa i?eeia? o/anoue o
caaacia/aii? oi/iino? a?aeai?o ai?iiaoeee-o?IE ia A-naeo? ?eainiie.

I?ioa aia?ao a?ineioaco a?eea c ?ioeo aoea??ioe/ieo i?aai?ci?a
ooieoe?iio? o a?aenooiino? ae??aeaeaeiaiai EF-3 aai eiai ?ic/eiiiai
aiaeiao. Iniaeeaee ?ioa?an o oeueiio ieai? noaiiaeyoue ?eainiie c ?ioeo
ieae/eo aoea??io, Tetrahymena pyriformis, ye? aieiae?thoue i?oeii
ca’ycaiith AO?/GOP-aciith aeoeai?noth, aeecueeith ca aeayeeie
ia?aiao?aie aei ae??aeaeaeiai? EF-3 AO?ace (Miyazaki, 1990).

Ui noino?oueny AO?acii? aeoeaiino? ?eainii aeueo aoea??io, oi a
e?oa?aoo?? c’yaeeeny ie?ai? aeac?aee ia ?? iiaeeeao iayai?noue, oi/a aei
ii/aoeo oe??? ?iaioe aiie aeeeeeaee aaeeeee noii?a ? ieoaiiy caeeoaeiny
a?aee?eoei.

Iaoa ?iaioe oa caaea/? aeine?aeaeaiue. Iaoith ?iaioe ? c’ynoaaiiy
ieoaiiy, /e iathoue aeniei i/euai? ?eainiie aeueo aoea??io caeaoi?noue
a?ae?ie?coaaoe AO?, oa ii??aiyiiy aeanoeainoae AO?ace c ae??aeaeaeiaei
EF-3 o ?ac? ?? aeyaeaiiy.

Aeey aeinyaiaiiy iinoaaeaii? iaoe ne?ae aoei ae??oeoe oae? ieoaiiy:

— aeine?aeeoe caeaoi?noue aeniei i/euaieo ?eainii c ?yaeo aeueo aoea??io
a?ae?ie?coaaoe AO?;

— ca iayaiino? ?eainiiii? AO?ace aea/eoe ?? aeanoeaino?, o oiio /ene?
noano?aoio niaoeeo?/i?noue, e?iaoe/i? ia?aiao?e ?aaeoe?? a?ae?ie?co AO?,
aieea ia ooieoe?iioaaiiy A-naeoo ?eainiie, /ooeea?noue aei ?ia?a?oi??a
EF-3;

— aecia/eoe, /e caeaoi? ?eainiie aeueo aoea??io noeioethaaoe
AO?/GOP-acio aeoeai?noue ae??aeaeaeiaiai EF-3.

Iaoeiaa iiaecia ?iaioe. Aia?oa aeieacaii, ui aeniei i/euai? ?eainiie
aeueo aoea??io i?noyoue i?oeii ca’ycaio AO?aco. Iiiaae 50 % AO?acii?
aeoeaiino? caa??aa?oueny o ?eainiiao iaa?oue i?ney a?aeieaaiiy 1,5 I
LiCl, ? o?eueee ?o ia?iaea 4 I LiCl, yea niieo/aia c ao?aoith /anoeie
?eainiiieo a?ee?a, i?ecaiaeeoue aei iaeaea iiaiiai cieeiaiiy caeaoiino?
?eainii a?ae?ie?coaaoe AO?. Iieacaii, ui AO?acia aeoeai?noue ?eainii ia
? iane?aeeii ?o caa?oaeiaiiy oaeoi?aie aeiiaaoe??.

Aia?oa i?iaaaeaii ii??aiyiiy ?eainiiii? c ae??aeaeaeiaei EF-3 ? aeyaeaii
?yae caaaeueieo aeanoeainoae, o oiio /ene? oe?ieo noano?aoio
niaoeeo?/i?noue oa i?eai?/aiiy niaoeeo?/ieie ?ia?a?oi?aie.

Aea/aii e?iaoe/i? ia?aiao?e ?eainiiii? AO?ace ia/?iee e?iey, iieacaii
iayai?noue aeaio aeoeaieo oeaio??a oa aeyaeaii noeioethth/o ae?th
ai?iiaoeee-o?IE ia eaoae?oe/io aeoeai?noue iaeiiai c ieo.

Aia?oa io?eiaii aeai?, ui na?ae/aoue i?i o/anoue ?eainiiii? AO?ace o
eiio?ie? iaae ooieoe?iioaaiiyi A-naeoa ?eainiie, aea a?aeaoaa?oueny
a?aea?? eiaeii-niaoeeo?/ieo o?IE. Anoaiiaeaii, ui ?eainiie aeueo
aoea??io, iath/e aeanio AO?acio aeoeai?noue, caa?aaee o i?ioean?
aaiethoe?? i?aeneethth/ee aeaiaio(e), iaiao?aeiee aeey noeioeyoe?? EF-3.

Oai?aoe/ia cia/aiiy ?iaioe. ?iaioa iineoue ooiaeaiaioaeueiee oa?aeoa? ?
?? oe?ii?noue iieyaa? o iiaiio c?icoi?ii? aeaoaeueieo iaoai?ci?a oeeeeo
aeiiaaoe?? iie?iaioeaeiiai eaioethao ia 80S ?eainii? oa aeyaeaii? iiaeo
iiaeeeaeo oi/ie i?eeeaaeaiiy oaeoi??a, ye? ?aaoeththoue a?ineioac a?eeo
o aeueo aoea??io.

Ca’ycie ?iaioe c iaoeiaeie i?ia?aiaie, ieaiaie, oaiaie. ?iaioa
aeeiioaaeany a ?aieao ieaio iaoeiai-aeine?aeieo ?ia?o a?aeae?eo
iaoai?ci?a o?aineyoe?? aaiaoe/ii? ?ioi?iaoe?? ?inoeoooo iieaeoey?ii?
a?ieia?? oa aaiaoeee IAI Oe?a?ie ca a?aioii ? 5.3/16 Aea?aeaaiiai oiiaeo
ooiaeaiaioaeueieo aeine?aeaeaiue ii oai? «C’ynoaaiiy ?ie? AO?acii?
aeoeaiino? ?eainii o a?ineioac? a?eea a aoea??io» oa ca a?aioii UBA 200
I?aeia?iaeiiai Ni?in?anueeiai oiiaeo.

Iniaenoee aianie caeiaoaa/a. ?iaioo aeeiiaii i?ae ea??aieoeoaii
ae.a.i., aeaaeai?ea A.A.?euenueei? o ni?a?ia?oieoeoa? c e.a.i. A.A.
Ia/a?aiei oa e.a.i. I.A. ?iaei?iith, c yeeie aoei iioae?eiaaii ni?euei?
?iaioe. Iniaenoei aianeii aaoi?a aeena?oaoe?? aoea o/anoue o ieaioaaii?
?iaioe, aeeiiaii? aenia?eiaio?a oa iaaiai?aii? iaea?aeaieo ?acoeueoao?a.
Aenia?eiaioaeuei? aeai?, iaaaaeai? a aeena?oaoe??, io?eiaii ca
aaciina?aaeiuei? o/ano? aaoi?a.

No?oeoo?a oa ianya ?iaioe. Aeena?oaoe?y neeaaea?oueny c anooio, iaeyaeo
e?oa?aoo?e, aenia?eiaioaeueii? /anoeie, yea aeeth/a? iien iaoa??ae?a oa
iaoiae?a aeine?aeaeaiiy, aeeeaae ?acoeueoao?a, ?o iaaiai?aiiy, aeniiae?a
oa nieneo oeeoiaaii? e?oa?aoo?e, yeee aeeth/a? 191 iaeiaioaaiiy. ?iaioo
aeeeaaeaii ia 107 noi??ieao, aeeth/ath/e 17 ?enoie?a oa 2 oaaeeoe?.

Ai?iaaoe?y ?iaioe. Iaoa??aee ?iaioe aeiiia?aeaeeny ia i?aeia?iaei?e
eiioa?aioe?? «Conference on the translational apparatus» (Berlin,
Germany, 1992).

Ioae?eaoe??. Ca iaoa??aeaie aeena?oaoe?? iioae?eiaaii 3 ?iaioe o
a?o/eciyieo oa caei?aeiiieo aeo?iaeao.

Iaoa??aee oa iaoiaee

O??n, iie? (U), NTP, GDP, AeOO ae?iaieoeoaa «Calbiochem», NOA.
2-ia?eaioiaoaiie, aeecien?oieao iao??th, MgCl2 — «Merk», ae?eeai?ae,
a?nae?eeai?ae, TEMED, io?ii?oeei, eoian? nei?e R-250 — «Serva», O?I.
Naoaaeaene, naoa?ice — «Pharmacia», Oaaoe?y. Neioaoe/i? iin??: DEAE-,
CM-ToyoPerl, — «Toyo-TCM», ssiii?y. A?ae?ien?aiaoeo — «Bio-Rad», NOA.
O?eueo?e «Whatman», Aaeeeia?eoai?y, «Millipore», NOA, «Sartorius-GmbH»,
O?I.

Iniiaiei ia’?eoii aeine?aeaeaiue aoea ia/?iea e?iey.

Io?eiaiiy ?eainiiieo noa/anoie i?iaiaeeee c aeoeaieo iie?nii ca iaoiaeii
(Falvey, 1970) o iiaeeo?eaoe?? (Iioaiia, 1986). ?o eiioeaio?aoe?th
aecia/aee, i?eioneath/e, ui 1 iae. A260/ie a?aeiia?aea? 28 iiieue/ie
aeey 40S noa/anoie oa 55 iiieue/ie aeey 60S noa/anoie. 80S ?eainiie
io?eioaaee ?aanioe?aoe??th 40S oa 60S noa/anoie. «Run-off» ?eainiie c
e?iey/i? oa ae/a/i? ia/?iee, a oaeiae ?aoeeoeioeeo?a e?iey, io?eioaaee c
iie?nii oeueo?aoeaio?eooaoaaiiyi o i?enooiino? 10 iI MgCL2I.

I?aia?ao noia?ieo A?Nac (EO 6.1.1) io?eioaaee a?aeiia?aeii (Von der
Haar, 1974). A?eeia? oaeoi?e aeiiaaoe?? (EF-1H oa EF-1a) io?eioaaee
nooi?i/anoei inaaeaeaiiyi iinoiie?niiaeueiiai noia?iaoaioo noeueoaoii
aiii?th c iine?aeoth/ith aaeue-o?eueo?aoe??th oa na???th o?iiaoia?ao?e
ia ?iiiiai?iieeao oa a?ae?ien?aiaoeo?. Io?eiaiiy oa i/euaiiy EF-3 c
ae??aeaeae?a i?iaiaeeee ye iienaii a (Skogerson, 1979).

Noia?io o?IE io?eioaaee ca iaoiaeii oaiieueii? aeno?aeoe?? c
iine?aeoth/ith o?iiaoia?ao??th ia DEAE-oeaetheic?,i/euaio o?IEPhe
-o?iiaoia?ao??th ia BD-oeaetheic?. Aeey io?eiaiiy [14N]Phe-o?IEPhe
cae?enithaaee i?aia?aoeaia ai?iiaoeeethaaiiy o?IEPhe c iiaeaeueoith
o?iiaoia?ao??th ia AAe-oeaetheic?. Nooi?iue caaaa/aiiy io?eiaieo
i?aia?ao?a neeaaeaea 1600 iiieue/iae. A260. N-aoeaoee-[14C]Phe-o?IEPhe
(AcPhe-o?IEPhe — aiaeia iaioeae?e-o?IE) io?eioaaee ca iaoiaeeeith
(Rappoport, 1974). Ca’ycoaaiiy Ac[14C]Phe-o?IEPhe c 80S ?eainiiaie
aei??thaaee ca aeiiiiiaith iaoiaeo o?eueo?aoe?? /a?ac i?o?ioeaetheici?
iaia?aie. Caiiaiaiiy ?-naeoo eiio?iethaaee ca aeiiiiiaith io?ii?oeeiiai?
?aaeoe?? (Rodnina, 1989). Oaeoi?caeaaeia ca’ycoaaiiy o?IE c ?eainiiith
i?iaiaeeee ye iienaii a ?iaio? (Rodnina, 1995).

Aecia/aiiy ATP/GPTacii? aeoeaiino? i?iaiaeeee ca iaoiaeii (Parmeggiani,
1981). ?aae?iaeoeai?noue i?ia aecia/aee o noeeioeeyoe?eiiio e?/eeueieeo.

Aeaeo?ioi?ac a?ee?a i?iaiaeeee ca (Laemmli, 1970) o i?enooiino? 0,1 %
aeiaeaoeeenoeueoaoo iao??th a 12,5% IAAA.

Aeey iienaiiy iae?i?eieo a?ao?e?a (oa ae?eyiie a?ao?e?a) o eii?aeeiaoao
?ae?-Oiono? aeei?enoiaoaaee aii??e/iee i?aeo?ae O?eea (Eo?aaiia, 1978).

A?ao?/iee aiae?c, ?ioa?iieyoe?th oa ?ic?aooiie e?iaoe/ieo ia?aiao??a
i?iaiaeeee ca aeiiiiiaith noaoenoe/ii? i?ia?aie «AsEasy» oa
niaoe?ae?ciaaiiai iaeaoo i?ia?ai aiae?co a?io?i?/ieo i?ioean?a
«Enzefitter».

?acoeueoaoe

Caeaoi?noue 80S ?eainii aeueo aoea??io a?ae?ie?coaaoe AO?

Iniiaiei caaaeaiiyi oeueiai ?icae?eo ?iaioe aoei c’ynoaaiiy ieoaiiy, /e
? o neeaae? 80S ?eainii aeueo aoea??io iiaeeeaee aiaeia oaeoi?o EF-3.
Aeey oeueiai aoei aea/aii caeaoi?noue 80S ?eainii ??ciiai iioiaeaeaiiy
a?ae?ie?coaaoe AO? oa GTP (?en. 1).

?en. 1. E?iaoeea a?ae?ie?co ATP/GTP «run-off» ?eainiiaie aeueo aoea??io:
?c ia/?iee aeea; ?c ia/?iee uo??a; ?c ?aoeeoeioeeo?a e?iey.

?aaeoe?y i?io?ea? aeineoue aoaeoeaii o i?enooiino? an?o i?aia?ao?a
?eainii — 10-20 iieaeoe AO?/?eainiia, oa. Ia ?en. 2 iieacaii, ui 80S
?eainiie, ?aeiino?oeiaai? c 40S oa 60S noaiaeeieoeue, oaeiae caeaoi?
a?ae?ie?coaaoe AO? aac aoaeue-yeiai aeiaeaoeiaiai ?ic/eiiiai a?eeiaiai
oaeoi?o.

?en. 2. E?iaoeea a?ae?ie?co ATP/GTP ?aanioe?eiaaieie 80S ?eainiiaie ?c
ia/?iee e?iey

Aeyaeeiny, ui GTP oaeiae a?ae?ie?co?oueny ye «run-off», oae ?
?aeiino?oeiaaieie 80S ?eainiiaie, oi/a e iaio aoaeoeaii, i?ae AO?.

I?iieaaiiy ?eainii 0,66 I (KCl + NH4Cl), ia?iaea iecueeeie
eiioeaio?aoe?yie Mg2+, oeaio?eooaoaaiiy o a?aae??io? naoa?ice,
aeenioe?aoe?y ?eainii oa ?aanioe?aoe?y ?o c noaiaeeieoeue ia
i?ecaiaeyoue aei cieeiaiiy AO?acii? aeoeaiino?. E??i oiai, ?eainiiaeueia
ATP/GTPacia aeoeai?noue ia i?eai?/o?oueny ooneaeiaith eeneioith oa
aioeo?eaie i?ioe EF-1 , ui na?ae/eoue i?ioe iia’ycaiiy oe??? aeoeaiino?
c aeii?oeaie oaeoi??a aeiiaaoe?? o ?eainiiieo i?aia?aoao.

Uia aeine?aeeoe, ye i?oeii iia’ycaia AO?acia aeoeai?noue c ?eainiiith,
inoaiith i?iieaaee c?inoath/eie eiioeaio?aoe?yie LiCl. sse aeaeii c ?en.
3, ?eainiie c ae/a/i? ia/?iee oa ?aoeeoeioeeo?a e?iey, i?iieo? 1 I oa
1,5 I LiCl, caa??aathoue 80 % oa 50 % ii/aoeiai? AO?acii? aeoeaiino?
a?aeiia?aeii, oi/a ao?a/athoue i?e oeueiio 55 % oa 78 % aeoeaiino? o
neioac? iie?oai?eaeai?io. O?eueee ia?iaea 4 I LiCl i?eai?/o? AO?acio
aeoeai?noue i?aeoe/ii iiai?noth. Oe? ?acoeueoaoe ua ?ac
i?aeoaa?aeaeothoue, ui AO?aca ? iaa?ae’?iiith naia 80S ?eainii.

?en. 3. E?iaoeea a?ae?ie?co ATP «run-off» ?eainiiaie c ia/?iee aeea oa
?aoeeoeioeeo?a e?iey.

Aeanoeaino? AO?ace 80S ?eainii ia/?iee e?iey

A ia?oeo aenia?eiaioao aoei aea/aii aieea iaeii- oa aeaioaaeaioieo
eao?ii?a ia ?eainiiio AO?aco. Aeoeai?noue oa?iaioo caeaaeeoue a?ae iaio
aeae?a eao?ii?a, i?e/iio ?o iioeiaeuei? eiioeaio?aoe?? ei?aeththoue c
oaeeie aeey iie?iaioeaeiiai neioaco.

E?iaoe/i? ia?aiao?e ?eainiiii? AO?ace aoei aea/aii o a?aeiia?aeiino? c
??aiyiiyi I?oaae?na-Iaioai. A?ao?/iee aiae?c o eii?aeeiaoao ?ae?-Oiono?
iieacaa iayai?noue aeaio e?iaoe/ii ??cieo eiiiiiaio, ui na?ae/eoue i?i
iayai?noue aeaio ??cieo naeo?a. Eiii’thoa?iee aiae?c aeicaieea
?icae?eeoe neeaaeia? oa ?ic?aooaaoe e?iaoe/i? ia?aiao?e aeey eiaeiiai
naeoo (oaae. 1).

Oaaeeoey 1

E?iaoe/i? ia?aiao?e AO?ace 80S ?eainii c ia/?iee e?iey

Aa-o?IEEIEcatieiieueoa.-1-Phe-o?IEPheI8711,5II101033,4+Phe-o?IEPheI8310,
8II161098,7

Eaoae?oe/i? aeanoeaino? aeaio naeo?a ?eainiiii? AO?ace nooo?ai
a?ae??ciythoueny. Aaaeeeai a?aecia/eoe, ui aeiaeaaaiiy Phe-o?IE neeueii
noeioeth? aeoeai?noue eeoa iaeiiai naeoo.

Nooo?aith iiae?ai?noth ?eainiiii? AO?ace c EF-3 aeyaeeiny ?ia?aoaaiiy ??
aeoeaiino? aaiaaeaoii aiii?th, yeee iaeaaeeoue aei eeane/ieo ?ia?a?oi??a
AO?ac i?ioiiieo ianin?a (?en. 4A). ?ioei aoaeoeaiei ?ia?a?oi?ii
?eainiiii? AO?ace aeyaeany aeeaei?ae ai?oei (?en. 4A).

?en. 4. Aieea aaiaaeaoa aiii?y oa ai?oeia ia AO?acio aeoeai?noue 80S
?eainii ?c ia/?iee e?iey.

?ia?a?oi?iee aiae?c iieacaa, ui iiae?aii EF-3 AO?aca 80S ?eainii ia?
oe?ieo noano?aoio niaoeeo?/i?noue (?en. 5).

?en. 5. ?ia?aoaaiiy ?eainiiii? AO?ace o i?enooiino? NDP, NTP oa ?o
aiaeia?a.

Aiaeiae AO? oa GTP, ui ia a?ae?ie?cothoueny, aeyaeeeny iaea?eueo
neeueieie ?ia?a?oi?aie, aoaeoeai?noue NTP ?ia?a?oi??a ciaioo?oueny a
?yaeo ITP, GTP, CTP/UTP; ADP oa GDP iathoue nooo?ai a?eueo iecueeo
ao?ii?noue aei aeoeaieo oeaio??a AO?ace.

Iecueea ao?ii?noue AO?ace aei NDP aeaco? ia oa, ui aeenioe?aoe?y
i?iaeoeoo ?aaeoe?? ia ? oaeaee?noue-e?i?ooth/ith noaae??th. Oie oaeo, ui
aiaeiae AO? oa GTP, ui ia a?ae?ie?cothoueny, iathoue a?eueoee
?ia?aoth/ee aieea, i?ae iiaeia aoei /aeaoe ia i?aenoaa? aea?iiey?iiai
ni?aa?aeiioaiiy, aeicaiey? i?eionoeoe, ui ca’ycaiee aiaeia aeenioe?th?
aeoaea iia?eueii ? ui ?eainiia iiaea aooe iei aeieiaaia o iaaiiio noai?.

Iiaeeeaa o/anoue ?eainiiii? AO?aceo ca’ycoaaii? o?IE c A-naeoii ?eainiie

Uia iaaeeceoeny aei ?icoi?iiy ooieoe?iiaeueii? ?ie? ?eainiiii? AO?ace,
aoei aea/aii aieea ??cieo oi?i o?IE ia ?? aeoeai?noue (oaae. 2).
Aeiaeaaaiiy «noncognate» o?IE aai iie?(U) ia aieeaa? ia ??aaiue
a?ae?ie?co AO?, a oie /an ye eiaeii-niaoeeo?/ia o?IEPhe iineeth? AO?acio
aeoeai?noue. Ua a?eueo ae?aciee noeioethth/ee aoaeo ciaeaeaii aeey
Phe-tRNAPhe oa AcPhe-tRNAPhe.

Aaaeeeai i?aee?aneeoe, ui ao?ii?noue oeeo oi?i o?IE aei A-naeoo ?eainiie
ia ii?yaeie aeua o ii??aiyii? c o?IE, a oie /an ye a?aeiinia
nii??aeiai?noue aei ?-naeoo ??cieo ooieoe?iiaeueieo oi?i o?IE ia?
cai?ioi?e oa?aeoa?. Ioaea, aaeeea aoaeoeai?noue ca?yaeaeaieo oi?i o?IE o
iineeaii? a?ae?ie?co AO? ii??aiyii c aeaaoeeeueiaaiith o?IE iiaea aooe
iia’ycaia c oei, ui noeioethth/ee aoaeo caeaaeeoue a?ae ca’ycoaaiiy o?IE
c A-naeoii ?eainiie.

Oaaeeoey 2

Aieea ooieoe?iiaeueieo oi?i o?IE ia AO?acio aeoeai?noue 80 S ?eainii ?
?eainiiieo noa/anoeoe ?c ia/?iee e?iey

??aaiue a?ae?ie?co AO?,%eiio?ieue100o?IE-Phe1,2 ieI100o?IEPhe0,6
ieI112o?IEPhe1,2 ieI 141Pha-o?IEPhe0,6 ieI154AcPha-o?IEPhe0,6 ieI 160

Oea i?eiouaiiy aoei ia?aa??aii a aenia?eiaioao, oeiiaee i?eeeaae yeeo
iaaaaeaii ia ?en. 6.

?en. 6. Aoaeo ca’ycoaaiiy AcPhe-tRNAPhe c A ? ? naeoaie ia AO?acio
aeoeai?noue.

sse aeaeii, caiiaiaiiy ?-naeoo AcPhe-tRNAPhe ia aieeaa? ia ??aaiue
a?ae?ie?co AO?, a oie /an ye e?ea?, ui a?aeiia?aeathoue ca?eueoaiith
AO?acii? aeoeaiino? oa ca’ycoaaiith noano?aoo c A-naeoii, iaeaea
ni?aiaaeathoue. Ioaea, ca’ycoaaiiy o?IE naia c A-naeoii ?eainiie ?
nooo?aei aeey ?? noeioethth/iai aoaeoo.

Aeieea? ieoaiiy, /e nooo?aa aeoeai?noue AO?ace aeey caiiaiaiiy A-naeoa
?eainiie, ye o aeiaaeeo c EF-3. Aea/aiiy aieeao aiaeiaa AO?, ui ia
a?ae?ie?co?oueny, aeyaeei iaeaea iiaio i?eai?/ai?noue EF-1 caeaaeiiai
ca’ycoaaiiy Phe-tRNAPhe c A-naeoii ?eainiie (?en. 7), oiaoi i?e
i?aeoe/ii iiaiiio i?eai?/aii? ?eainiiii? AO?ace A-naeo ? aeieiaaiei.

?en. 7. Aieea AdoPP[NH]P ia ??aaiue EF-1-caeaaeiiai ca’ycoaaiiy
Phe-tRNAPhe c A-naeoii.

Oaeei /eiii, iaaaaeai? o aeaiiio ?icae?e? ?acoeueoaoe aeathoue i?aenoaao
aaaaeaoe, ui ?eainiiia AO?aca aa?a o/anoue o eiio?ie? ooieoe?iioaaiiy
A-naeoa iiae?aii ae??aeaeaeiaiio EF-3.

Noeioeyoe?y aeoeaiino? ae??aeaeaeiaiai EF-3?eainiiaie ia/?iee e?iey

O iiaeaeueoeo aenia?eiaioao aea/aeanue iiaeeea?noue noeioeyoe??
ae??aeaeaeiaiai EF-3 ?eainiiaie c aeueo aoea??io. Anoaiiaeaii, ui i?e
iaaieo oiiaao caaaeueia AO?acia aeoeai?noue cia/ii ia?aa?eueoo? noio
aeoeaiinoae ?eainii oa EF-3, oi/a e?iaoeea AO? a?ae?ie?co cia/ii
iia?euei?oa o aaoa?ieia?/i?e nenoai? (?en. 8A).

?en. 8. Caeaaei?noue AO? a?ae?ie?co a?ae /ano oa eiioeaio?aoe?? AO?.

Oe?eaai, ui e?eaa caeaaeiino? oaeaeeino? a?ae?ie?co a?ae eiioeaio?aoe??
AO? o i?enooiino? EF-3 oa 80S ?eainii ia ia? oi?ie a?ia?aiee, ye oea
oa?aeoa?ii aeey ?eainiiii? AO?ace (?en. 8A). ?en. 9 aeaiiino?o? ??cio
caeaaei?noue ??aiy noeioeyoe?? a?ae eiioeaio?aoe?? EF-3 oa ?eainii ?c
ia/?iee e?iey aai ae??aeaeaeiaeo.

?en. 9. Caeaaei?noue ??aith AO? a?ae?ie?co a?ae eiioeaio?aoe??
ae??aeaeaeiaiai EF-3 aai ?eainii.

Iaaaaeai? aeai? aeacothoue aeacothoue ia oa, ui aoaeo noeioeyoe??
iaoiiaeaiee ca?eueoaiiyi aeoeaiino? EF-3, a ia ?eainiiii? AO?ace. Oeae
aeniiaie aoei i?aeoaa?aeaeaii a aenia?eiaioao c EF-3, i?a?ieoaaiei ?c
aioeo?eaie i?ioe iueiai aeey ?ia?aoaaiiy eiai AO?acii? aeoeaiino?. Oaeei
/eiii, ?eainiiiee i?aeneethth/ee (?aaoeyoi?iee?) aeaiaio aeey AO?ace
ae??aeaeaeiaiai EF-3 caa??any a i?ioean? aaiethoe??, ? eiai o/anoue o
ooieoe?iioaaii? aeania ?eainiiii? AO?ace aeueo aoea??io yaey? niaith
iniaeeaee ?ioa?an aeey iiaeaeueoeo aeine?aeaeaiue.

Caeeth/aiiy

A?aee?eooy ?eainiiii? AO?ace Tetrahymena iinoaaeei caieoaiiy i?i
i?eioeeiiao oi?eaeuei?noue ae??aeaeaeiaiai EF-3 oa i?i iayai?noue
?eainiiii? AO?ace o aeueo aoea??io. ?acoeueoaoe, ui ie io?eiaee,
iaeiicia/ii na?ae/aoue i?i oa, ui ?eainiie c ?yaeo aeueo aoea??io
i?noyoue i?oeii ca’ycaio ATP/GTPacio aeoeai?noue. sse a?aeiii,
aoea??ioe/ia ?eainiia ia i?noeoue a?eeo, yeee a?aeiia?aea? ca ?ici??aie
EF-3 (115 kDa). I?ioa ia aeeeth/aii, ui ?eainiiiee aiaeia EF-3
neeaaea?oueny c aeaio iie?iaioeaeieo eaioetha?a aai ui ATPacio
aeoeai?noue iathoue a?eee, ni?euei? aeey aeaio ?eainiiieo noaiaeeieoeue
? ?ici?uai? i?ae ieie.

Anoaiiaeaii i?eiaeii? o?e aeanoeaino?, ni?euei? aeey AO?ace ?eainii
ia/?iee e?iey oa ae??aeaeaeiaiai EF-3, ye? iathoue i?eioeeiiai aaaeeeaee
oa?aeoa?: 1) oe?iea noano?aoia niaoeeo?/i?noue, 2) ?ia?aoaaiiy
aaiaaeaoii aiii?th, yeee ? eeane/iei ?ia?a?oi?ii AO?ac i?ioiiieo
ianin?a, 3) aeaa aeoeai? oeaio?e ?eainiiii? AO?ace a?aeiia?aeathoue
iayaiino? aeaio AO? ca’ycoth/eo ae?eyiie o no?oeoo?? ae??aeaeaeiaiai
EF-3.

Io?eiai? aeai? aeicaieythoue i?eionoeoe, ui ?eainiiia AO?aca aeueo
aoea??io aa?a o/anoue o eiio?ie? ooieoe?iioaaiiy A-naeoo, aea
a?aeaoaa?oueny a?aea?? eiaeii-niaoeeo?/ieo o?IE. A oie aea /an a?aeiii,
ui ?nio? iaaia eii?aeeiaoe?y i?ae A- oa A-naeoaie 80 S ?eainii.

Aaaeeeaith ? oaeiae aecia/aia a ?iaio? caeaoi?noue 80 S ?eainii, ui
iathoue aeanio AO?acio aeoeai?noue, noeioethaaoe ae??aeaeaeiaee EF-3,
oiaoi caa?aaeaiiy a i?ioean? aaiethoe?? ?aaoeyoi?iiai aeaiaioo(?a).

aeniiaee

1. Anoaiiaeaii, ui ye «run-off» ?eainiie, oae ? ?aeiino?oeiaai? c
noaiaeeieoeue 80S ?eainiie oaa?eiiiai iioiaeaeaiiy, iathoue AO?/GTPacio
aeoeai?noue, yea cieea? o?eueee i?ney i?iieaaiiy ?eainii 4 I LiCl, ui
niieo/aii c ao?aoith cia/ii? /anoeie ?eainiiieo a?ee?a.

2. Iieacaii, ui iiae?aii AO?ac? ae??aeaeaeiaiai EF-3 ?eainiiia AO?aca
ia/?iee e?iey oa?aeoa?eco?oueny oe?ieith noano?aoiith niaoeeo?/i?noth,
i?e/iio aoaeoeai?noue NTP ?ia?a?oi??a a?ae?ie?co AO? ciaioo?oueny a
?yaeo ITP, GTP, CTP/UTP.

3. E?iaoe/iee aiae?c iieacaa, ui AO?aca 80S ?eainii ia? aeaa aeoeai?
oeaio?e, ye? a?ae??ciythoueny ca cia/aiiyie EI oa Ecat. Eaoae?oe/ia
aeoeai?noue iaeiiai c oeaio??a nooo?ai i?aeaeuo?oueny o i?enooiino?
eiaeii-niaoeeo?/ii? ai?iiaoeee-o?IE, o oie /an ye ?ioee oeaio? ia
?aaoeth?oueny o?IE.

4. Iieacaia o/anoue ?eainiiii? AO?ace aeueo aoea??io o eiio?ie? iaae
ooieoe?iioaaiiyi A-naeoo ?eainiie, aea a?aeaoaa?oueny a?aea??
eiaeii-niaoeeo?/ieo o?IE i?ae /an iie?iaioeaeiiai neioaco. I?i oea
na?ae/eoue noeioeyoe?y aeoeaiino? ?eainiiii? AO?ace i?e ca’ycoaaii?
ai?iiaoeee- /e iaioeae?e-o?IE c A-naeoii ?eainiie oa aeieoaaiiy A-naeoa
aeey EF-1-caeaaeiiai ca’ycoaaiiy ai?iiaoeee-o?IE i?e i?eai?/aii?
AO?acii? aeoeaiino? aiaeiaii AO?, ui ia a?ae?ie?co?oueny.

5. Anoaiiaeaii, ui 80S ?eainiie ia/?iee e?iey caeaoi? nooo?ai
noeioethaaoe AO?acio aeoeai?noue ae??aeaeaeiaiai EF-3, ui na?ae/eoue i?i
caa?aaeaiiy ia ?eainii? a i?ioean? aaiethoe?? ye ae?eyiee ca’ycoaaiiy
EF-3, oae ? i?aeneethth/iai aeaiaioo.

NIENIE I?AOeUe IIOAE?EIAAIEO CA OAIITH AeENA?OAOe??

1. Marina V. Rodnina, Alexander I. Serebryanik, Galina V. Ovcharenko
and Anna V. El’skaya. ATPase strongly bound to higher eukaryotic
ribosomes. // Eur. J. Biochem. — 1994. — 22, — P. 305-310.

2. A.V. El’skaya, A.I. Serebryanik, G.V. Ovcharenko. Stimulation of
yeast EF-3 factor by mammalian ribosomes. // Oe?. aeioei. aeo?i. — 1995,
o. 67, ? 6. — N. 28-32.

3. A.E. Na?aa?yiee, N.N, Iaeue/aaneee, A.A. Aeueneay. ?eainiiiay AO?aca
aunoeo yoea?eio. // IAI Oe?aeiu. Einoeooo iieaeoey?iie aeieiaee e
aaiaoeee. I?ai?eio. Eeaa 1997.

AIIOAOe?ss

Na?aa?yiee I.?. ?eainiiia AO?aca aeueo aoea??io. — ?oeiien.

Aeena?oaoe?y ia caeiaoooy iaoeiaiai nooiaiy eaiaeeaeaoa a?ieia?/ieo
iaoe ca niaoe?aeuei?noth 03.00.03 — iieaeoey?ia a?ieia?y. — ?inoeooo
iieaeoey?ii? a?ieia?? oa aaiaoeee IAI Oe?a?ie, Ee?a, 1998.

Aia?oa aeieacaii, ui aeniei i/euai? ?eainiie aeueo aoea??io i?noyoue
i?oeii ca’ ycaio AO?aco oa aea/aii ?? aeanoeaino?. Iieacaii iayai?noue
aeaio aeoeaieo oeaio??a oa aeyaeaii noeioethth/o ae?th ai?iiaoeee-o?IE
ia eaoae?oe/io aeoeai?noue iaeiiai c ieo. Io?eiaii aeai? i?i o/anoue
?eainiiii? AO?ace o eiio?ie? iaae ooieoe?iioaaiiyi A-naeoa ?eainiie, aea
a?aeaoaa?oueny a?aea?? eiaeii-niaoeeo?/ieo o?IE. I?iaaaeaii ii??aiyiiy
?eainiiii? AO?ace c ae??aeaeaeiaei EF-3 ? aeyaeaii ?yae caaaeueieo
aeanoeainoae. Anoaiiaeaii, ui ?eainiie aeueo aoea??io, iath/e aeanio
AO?acio aeoeai?noue, caa?aaee o i?ioean? aaiethoe?? i?aeneethth/ee
aeaiaio(e), iaiao?aeiee aeey noeioeyoe?? EF-3. Oai?aoe/ia cia/aiiy
?iaioe iieyaa? o iiaiio ?icoi?ii? aeaoaeueieo iaoai?ci?a oeeeeo
aeiiaaoe?? iie?iaioeaeiiai eaioethao ia 80S ?eainii? oa aeyaeaii? iiaeo
iiaeeeaeo oi/ie i?eeeaaeaiiy oaeoi??a, ye? ?aaoeththoue a?ineioac a?eeo
o aeueo aoea??io.

Eeth/ia? neiaa: noa/anoeie ?eainii, AO?acia aeoeai?noue, oeeee
aeiiaaoe??, EF-3 ae??aeaeae?a.

AIIIOAOeEss

Na?aa?yiee I.?. ?eainiiiay AO?aca aunoeo yoea?eio. —

?oeiienue.

Aeenna?oaoeey ia nieneaiea o/aiie noaiaie eaiaeeaeaoa aeieiae/aneeo
iaoe ii niaoeeaeueiinoe 03.00.03 — iieaeoey?iay aeieiaey. — Einoeooo
iieaeoey?iie aeieiaee e aaiaoeee IAI Oe?aeiu, Eeaa, 1998.

Aia?aua aeieacaii, /oi aeniei i/euaiiua ?eainiiu aunoeo yoea?eio
niaea?aeao i?i/ii naycaiioth AO?aco eeco/aiu aa naienoaa. Iieacaii
iaee/ea aeaoo aeoeaiuo oeaio?ia e auyaeaii noeioee?othuaa aeaenoaea
aieiiaoeee-o?IE ia eaoaeeoe/aneoth aeoeaiinoue iaeiiai ec ieo. Iieo/aiu
aeaiiua ia o/anoee ?eainiiiie AO?acu a eiio?iea iaae ooieoeeiie?iaaieai
A-naeoa ?eainiiu, aaea i?ienoiaeeo ioai? eiaeii-niaoeeoe/aneeo o?IE.
I?iaaaeaii n?aaiaiea ?eainiiiie AO?acu n ae?iaeaeaaui EF-3 e auyaeai
?yae iaueo naienoa. Onoaiiaeaii, /oi ?eainiiu aunoeo yoea?eio, eiay
nianoaaiioth AO?acioth aeoeaiinoue, nio?aieee a i?ioeanna yaiethoeee
oneeeoaeueiue yeaiaio(u), iaiaoiaeeiue aeey noeioeyoeee EF-3.
Oai?aoe/aneia cia/aiea ?aaiou ninoieo a iiaii iiieiaiee aeaoaeueiuo
iaoaieciia oeeeea yeiiaaoeee iieeiaioeaeiie oeaie ia 80S ?eainiia e
auyaeaiee? iiauo aiciiaeiuo oi/ae i?eeiaeaiey oaeoi?ia, eioi?ua
?aaoee?otho aeineioac aaeea o aunoeo yoea?eio.

Eeth/aaua neiaa: noa/anoeoeu ?eainii, AO?aciay aeoeaiinoue, oeeee
yeiiaaoeee, EF-3 ae?iaeaeae.

SUMMARY

A.I. SEREBRYANIK. The ribosomal ATPase from higher eukaryotes.

— Manuscript

Thesis for a degree of Doctor of Philisophy (PhD in Biology, Special
Option — Molecular Biology.- Institute of Molecular Biology and
Genetics, National Academy of Sciences of Ukraine, Kyiv, 1998.

It has been first shown that highly purified ribosomes from higher
eukaryotes are strongly associated with ATPase activity, and its
properties have been studied. Two active sites have been revealed, one
of them may be stimulated by aminoacyl-tRNA. The data obtained show
participation of the ATPase in the regulation of the ribosomal A-site
responsible for the selection of cognate tRNA. A comparative analysis of
the ribosomal ATPase and yeast EF-3 has elucidated some common
properties. Higher eukaryotic ribosomes, even though possessing
intrinsic ATPase, are found to preserve in the course of evolution an
enhancing element for the yeast EF-3 ATPase stimulation. The thesis
contributes to a new understanding of detailed mechanisms of the
elongation cicle on 80S ribosome, including the elucidation of novel
points of protein biosynthesis regulation in mammals.

Key words: ribosomal subunits, ATPase activity, the elongation cycle,
yeast EF-3.

Похожие записи