I1crnrny10 ,n_Jrn: o6pa3oBaHHH ,llJIMepoB II CBH3bIBaHHH c llHK [Gentz R. et al., 1989; Turner R., Tjian R., 1989].
BrrpycHaH npOTea3a C MOJieKyJrn:pHoii: Maccoil: 9000-11 000 naXO,ll_HTCH Ha N-KOHJJ,C npe,n_meCTBeHHHKa pol H OTllI.eTIJIHeTCH
OT npe,n_meCTBeHHHKa B pe3yJibTaTe ayTOKaTaJIHTWieCKOro npo I(ecca [Debouk C. ct al., 1987]. <l>yHKUHH ee 3aKJIIOIJaeTcH B
1rnpe3aHHH npe,n_ruecTBeHHHKOB gag II pol B cneumjrntrecKHX ylJaCTKaX. <l>yHKJJ,HH npoTea3bI cyrnecTneHHhI I-IC TOJibKO ,ll_JIH npoueccHI-Ira 6eJIKOB cep,n_ueBHHLI, HO n IlOlJKOBaI-IHH BHpyca
11 npoHB.nemrn ero HHq:>eKUHOHH0!"1 aKTHBHOCTH [Kohl N. et al., 1988; Peng C. et al., 1989]. MyTare11e3 n Ko;uipyromeil: npo
Tea3y o6JiaCTH npHBO,ll_HT K 06pa3onarrmo HeHHcpeKJJ,HOHHOrO nrrpyca, co,n_ep2Kam.ero npeJirnecTBCHHHK gag - pol, HMeromero cepJiuenrrHy KOJibJJ,enrr,n_Hoii: cpopMLI H notJKy1omeroc51 ne c KJie
TOIJHoil: nonepxHOCTH, a B HHTOTIJia3MaTHlJeCKHe BaKyOJIH [Peng C. et al., 1989].
TTpoTea3a npHHa,n_Jie2KHT K ceMei1cTBY acnapTHHOBbix no CJie}J,yIOIIJ:HM npH3HaKaM: 1) HaJIHlJHe IlOCJie}IOBaTCJibHOCTeH Acn -Tprr - fJIH B MOJieKyJie; 2) nOTepH cpepMeHTaTHBHOH aKTHBHOCTH nprr MyTaJJ,HHX, npHBOJIHllI.HX K 3aMeHe Acn Ha AJia; 3) THTIHIJHaH CTPYKTypa aKTHBHoro neHTpa, onpe,n_eJIHe 'v!aH C IlOMOll(bIO KOMilhlOTepHoro 3H3JIH33. TToJiyLJeHbI KpHCTaJI JJHlJeCKHe CTPYKTYP"' npOTea31,1, wiylleHrre KOTOpbIX noKa3aJio, lJTO npOTea3a 5IBJI51C'TC5I CHMMCTPIJT!HhIM }IHMepoM C aKTHBHbIM ueHTpOM, CXOJIHhIM C TaKOBbIM TICTICHHOTIO,ll_06HI,IX npoTea:i
(nencHH, peHHH, KaTencHHbI D II E) [Wlodawer A. et al., 1989]. TTp11 cpanHeHHH npoTea3bI Bl'Jl-I c npOTeaJaMH ,n_pyrnx
peTpOBHpycoB 6LIJIH o6Hapy2KCHbI 3H311HTCJTbHbie pa3JIWIH5!,
oco6eHHO B TOTIOJIOrHH o6JiacTH nonepxHocTel'I ,n_HMepoB [Blun dell T., Pearl L., 1989].
TTpoTea3a o6JI3}I3eT 3HTHreHHLIMH CBO ICTBaMH H BLI3b!Ba CT B 70% cJiyqaen y 3apa2Kem11,1x BHq mm 06pa3onaHHe 3HTIITCJT. AHTHTeJia npOTHB npoTC33bI IlO5IBJI51IOTCH JrHIIIbcny CTH 1-2 ro,n_a nocJie cepo1rn1rnepcHH, ex nnp napaCTaeT B TelfeHHe IlOCJie,n_yromHx JieT, a B HeKOTOpLIX CJiylJaHX OHH HC lJC3aIOT. Y 60JibHbIX CTTI1ll aHTilTeJia npOTHB IlpOTea3bl HC
o6Hapy2KHB3IOTCH, BO3MO)KHO, B CB5I3H C HX HC1Ie3HOBeHHeM Ha noJ,n_HIIX cTa,n_nHx 6oJie3HH [Boucher C. ct al., 1989]. TTocJie)I
Hee MO)KCT npomoihrr Jrn6o B peJyJibTaTe JIIIccpyHKUHH B-KJie TOK, JIH6o H3-33 cpopMHpOB3HH5I IlMMYHHblX KOMTIJieKCOB C BII PVCHOH npoTea30H npII 3cpcpeKTHBIIOI1 penJIIIKaJJ,IIH BIIpyca. TTpOTea3a npe,n_CTaBJIHeT co6ol'J: yn3BIIMYIO MnmeHb ,n_JIH xn 'v!HoTepaneBTJil1ec1rnro BMCllI3TC,TTbCTR3, pe3y,nhT8TOM KOToporo MO)KCT 6hITh IIH3KTIIB3HII5T cpepMeIIT3TIIBIIOJ'i :1KTHRJ-IOCTII H.THI 6JIOKHpoBaHH5I ,ll_HMepH3aJJ,HH MOJieKyJI.
npo,nyKTbl retta env HBJlH!0TC5I fJUIK0rrpoTeH..QaMH. 9m 6eJIKH urpaior 0CH0BHYIO poJib B 11arore1-1ese nmpeKunn: 06yc
.110BJIHBaIOT npHKpenneirne mipyca K KJ!CT0'lH0MY peuenropy lI np0HHKH0BCHHe B KJ1eTKy nyreM CJI!IHI-IIUI MeM6paH, Bbl3bIBaIOT 06pas0Ba1-rne cHHUHTHH 11 pacnpocrpaire1rne Bnpyca H3 KneTKH
B KJleTKY, a TaKlli.e Hapywemie cpyHKllJIH |
T-JIHM<:pOUHTOB |
(CM. HF!:l:Ke). 9TH )Ke 6eJ1Kll, 9KCTI0HttpoBaHIILie |
Ha TI0BepXH0CTH |
BHpH0Ha H 3apa)KeHHoii KJICTKll, c.1y}K3T J].OCTynHOH MHilleHbIO J].JHI HMMYHHOi'IaTaKH. O6a I"JlHKonpoTeH)la, gp120 (Ha)lMeM6paHHblH 6eJJOK) II gp4l (rpaHCMeM6paHHbIH 6e.,10K), o6pa3YIOTCH nprr 1rnpesamrn npemnecTBeHHHKa gpl60 H 0Tnn1ia10T cn OT II0BepxH0CTHblX 6e.'1KOB JJ.pyrllx perpoBIIpycoB 60JiblliH· MII pasMepaMll, llHTeHCllBHblMH rJ1IIK03H.111p0BaHllCM, IIlH!)OI<OH BapHa6eJibHOCTblO 3MllH0KIICJl0THhIX 0CTaTK0B. BeJIOK gpl 60 C0)lep2KHT 3 rtt)lpocpo6m,Ie o6naCTll'. CllfH3.'1bH35l TIOCJie,!I,0Ba TCv'lbH0CTb (OCT3TKH 1-30), N-KOHCIJ. gp41 (ocTaTKH 512541), o6JI3CTb B cepe,!I,HHe M0JieKyJlhl gp41 (ocTaTKII 684706). B speJI0M gp160 CHrHaJibHaH II0CJIC,IJ.OBaTeJibHOCTb OTpe saeTCH, H gp160 BCTpaHBaeTCH B J!IlillI)].HhIH 611cJIOH TIJIa3Ma TII1IeCKOH MCM6paHhl TaKHM o6pa30M, lJT0 Bech gpl20 (OCT3TKH 31-510) 11 IlOJl0BHHa gp41 c N-KOHUa. (oCTaTKH 512-683} H3X0J].HTC5I Ha IT0BepXH0CTH sapa)KeJIHOii KJICTKH. KoMIIbIOTep H A aHaJ1H3 H KaprHp0BaHIIe 9111IT0D0B IT03B0JllIJlH nonylJHTb M0,Il.CJlb opHenTaIJ.Hll f,1llK011pOTeH,!l.0B B e11pyc1-1oii H KJieTOl! HOH MeM6pattax '[Modrow S. et al., 1987; Berman P. et al., 1988]. B oTJrn1rne or rrpeJJ.lllecrBy1omeii M0Jl.e.1111, npeJI,rr0.TJa ra1omer"1 TpexKpan1oe 11epece11em1e rJ1HKonpoTeH)lOM MeM6pa Hb1 [Gallo R., 1986], 60,1ee npocTa5I M0,n:eJib npeJu10.1Jaraer C,ll,lIHCTI3eHHYIO TpaHcMeM6parrnyio O0JiaCTb, KOTOpoJi 6eJIOK «3aHK0pHBaCTC51>> B ,'1HTIHJJ.H0M 6HCJIOe, C00TBeTCTBYIOIUYI0 0CTaTK3M 684-706, a 0CTaTKH 707-856 B BHJJ.e rn,n,pocjJHJih
H0ro «XB0CTa>., rrpmrnKaIOT 11epe3 MCM6pa1-1y (pHC. 11) [Ber
man P. ct al., 1988].
,IVrn npoHBJiemrn cpyHKILlfll rJrnKonpoTCHA0B 6oJibIIIoe 3Ha qem1e HMCIOT HX opneuTall,H5I B JllIIIIl,ll,H0M 6HCJIOe BHpH0l!a II 3apmKeHH01I KJICTKH, H3Jll{lJlIC rn,npocpo6HblX Ii 3apa2KeHHhlX o6JiaCTCH B C0CTaBe M0JieKyJih[, 1rny11eH11e fJIHK03HJIIIp0Ba!-IIISI, qepe,ll,0B3HHC K0HCepBaTIIBHbJX H eap1rn6eJibHblX o6Jiacreii, Ha Jrn1rne I]JlCTeHHOBb!X ocTaTK0B lI ,np.
5eJIOK·I1pC}JJJ1CCTBCHHIIK gp160 co;i:ep)KIIT 861 aMIIHOKHC JIOTHblH 0CTaT0K, gp120 - 488 n gp41 - 343 0CTaTKa. B co CTaBe gpl20 I!MCCTCH 25 ytiaCTK0B rJIHK03HJIIIpoeaHHH, gp41- 5 yqac·rnoe; yrneB0AHbre ueno11KII coc1·asJ15IIOT no Macce 50%' M0JieKyJrnpHOH Macc11 6eJIKOB env, HaMH0f0 rrpeB0CX0)lH l!X'. K0JlJ{t!eCTB0 B rJlllK0IIp0Te11,nax JI.pyr1-1x 060jlO'!e1rnbIX BapycoB.
CJie)].0BclTeJiblI0CTH LJepeJJ.yI0TC51 c BapHa6eJibHblMH yLJaCTKaMll. KoncepBaTHBHbie yLJaCTKll B0BJICLJCHLI B KpH'rWieCKHe C06bITlI5I peIIJIHK3IJJIOHII0ro IJ)IKJJa: Y3IJaB3IIHC l)CIJ,CIIT0j)a, npoIJ,eCCHHl' M0JICKYJILI, co6LITil5I II0CJIC aJ].cop61I,HII BHpycHLIX LJaCTllll, Ha KJieTKc n JJ.p. Bapua6cJ1L11L1c Y'JaCTKII TaK2Kc mpamr Ba.tKHYIO po,'lb B HIHpCKIJ,lIOHHOM llj)0IJ,ecce, 0HH B3aHMO)].CHCTBYIOT C HeH rpaJIH3YI0lll,HMll aHTHTCJiaMH, BbIII0JIILHI0T J)}l)J, cj)yHKIJ,HH TI0CJie
a)\cop61I,HH |
BHpyca |
Ha KJieTKe. |
MoJ1cKyJia |
gp 120 |
co,ll.ep)IUIT |
6 K0HcepBaTHBHb!X |
Y'JaCTK0B, |
roM0JIOJ'l11 IHL1X y |
B114-1 lI |
BI14-2, u |
3 napHa6eJibHb!X Y'JaCTKa. )],Jrn |
K0HCepBaTHBHLIX |
yqaCTK0B HeO.tKH)].3lIHOH 5IBJl5IeTC5I npyKrypHa5I r0M0JI0fH5I lIX C K0HcepBaTHBHbIMII yLJaCTKaMH T5I.tKeJIOll IJ,em, HMMYH0rJI06y Jlllll0B LCordonnier A. et al., 1989]. O6JiacTb, yLJaCTBYIOW.a5I B CB5I3L!BaJHIII C peIJ,eIIT0poM, CJI0.tKII0 apaIDKHpoBaHa, ll aMHH0KlICJI0THLie II0CJie)].0BaTeJibH0CTll, Y3Ha!Olll,HC peIJ,enrop, no ,ll.aH HblM pa3HLIX aBT0p0B, BapbHpy10T. YKa3LIBaIOT TaKHe II0CJie ,ll.0BaTeJILH0CTlI, KaK 414-429, 427-432, 431-442, 487-499 lI Jl.j).
Tp11 Bap11a6eJILHLIX y,1acrKa B o6Jiacrn 128-, 267- H 308-ro aMHH0KHCJI0THLIX 0CTaTK0B Hrpa!0T Ba.tKHYI0 ))0J!b 13 npoIJ,ecce pcnJIHKaIJ,1111 BHpyca [Willey R_. ct al., Hl89J. B cocraue 3-fl Bapna6eJibHOH o6JiaCTll Haxo) HTC5I 0ClI0BHa.H )].0MlllI3HTa, B3aH MO,ll.CHCTBYIOlll,a5I c BHpycHc11TpaJm3y1ow.IIMHaHTHTCJiaMH [Ja vahcrian K. et al., 1989]. Bee rp11 yLJacrKa M0JICKYJib! B3aHM0- )].CHCTBYIOT ,LJ,pyr C JJ.pyroM, II 3T0 B3311MO)].eHCTBHe Ba)KlI0 )].Jl5I 3(j)(_peKTHBHOH npoJJ.yKIJ,HlI 13Hp110H0B. 3aMeHa 0)].H0ro aMHH0KHCJI0TH0ro ocrarKa BC)].CT K 6JJOKIIpouaHJIIO llll(j)CKIJ,IIOHHOro npoIJ,ecca [Willey R. et al., 1989; Felscr T. cl al., 1989].
Y TpaHcMeM6pammro 6eJIKa gp41 Ha1160Jree Ba.tKHLie ,ll.e TCpMIIHaHTbI - cneIJ,Hcjrn'leCKile Y'!aCTirn, B3aHMO,LJ,eHCTBYIOW.He C HCHTpaJIH3YIOW.HMll aHT!ITC,TaMH, pery,rnropHL!MlI 6eJIKaMH, CJIHBaIOlll,lIH IIeIITll)]., Y'I3CT0K, CB5I3bll3cl!O lll,HHC5I C gp120, Ha X0,ll.5ITC51 B noJioBHHe, 11pu:1craIOw.cflK N-KOHIJ,y. BcpwHHa Mo JieKyJibI 6orara 0CTaTKaMH ll,HCTClil-Ja, C0,il.Cj).tKlIT HMMYH0,ll.0MlI HaHTHYI0 o6JJaCTb H Y'JaCTKH rJIHK03HJIIIpoBaHH5I. BLI,ll.aIO lll,lIHC5I K0HeIJ, 5IBJ15ICTC5I CJIHBaIOlll,lIM neIITH)].0M H BKJIIOLiaeT rH)].pocj>o6HLIH IJ,eHrp LFLGFL, cxo,ll.HLIH c raK0BbIM cHrnaJib Horo neIITH,ll.a. IIerJieo6pa3Ha5I crpyKrypa, 6oraTa5I C H T (5 H3 11), B3aHMO)].eflcrnyer c gpl20. 3areM cJie,ll.yer HHTeH CHBH0 3apB.tKeHHa5I o6Jiacrb, cj>opMHpyiow.aBa-cnHpaJib, cra6HJIH3HpyeMy10 MH0.tKeCTBeHHLIMH napaMH H0H0B. BbICOK0rH,ll. pocj)o6Ha5I o6JiaCTL, rrp0HH3b!BaIOlll,a5I MeM6paHy, rrpe,ll.CT3BJieHa
a-c1rnpaJiblO C pe3KlIMH IIOB0poraMII BHTK0B II0CJie BLIX0,LJ,a II3 JIIIIIHJl.II0ro 6HcJI05I [Gallaher W. ct al., 1989] {cxeMa 2).
I-Ieo6bI'IHbIM CBOHCTBOM rpaHCMeM6pmrnoro 6eJIKa Bl14
IJOMOIUblO KOMilblOTCpHoro
IJ
lfaJIOMaHHhle Jll!HHH - a-c1111paJIH. I - CJIHBaIO ll.J.Hii nenTH.rr,; 2 - HMl\.1YHOJI.0MHH3HTHcl51 o6JiaCTb· J - 06JiaCTb CBll3bIBaHH}I C gpJ20; 4 - Tp,rn : MeM6paHaan o6JiaCTb; 5 - U.HTOIIJicl3MclTHlJ.CCI\Hii: yqacTOK; 6 - /(HCYJihcj,H/(Hal! CDll3b. CTpeJIKH - callThl rJIHK03HJI1ipoea1rna.
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HBJUieTCH HaJUiqIIe )].JIIIHH0ro l(HT0- |
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IlJia3MaTHqecK0ro XB0CTa ..:ha 06- |
|
JiaCTb |
C0J].ep.tKHT |
KOmpopMalI,IIOH |
|
Hbie )].eTepMIIHaHTbl II cueu_IIqmqc |
ae |
CKIIe |
II0CJICJ].0BaTeJibH0CTH, |
HMeIO |
|
IUIIe |
onpeAeJiettHoe |
3HaqettHe |
AJIH |
|
pacnpocTpattemrn m1pyca H npoHB |
|
JieHIIH |
lI,HT0IIaToreHH0CTH. |
|
TTpII |
|
n3yqeHIIH natteJIH MyTaHT0B BI1Y.-1 |
|
c J].eJielI,HHMH B o6JiaCTH 6-14-ro |
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aMIIH0KIICJI0TH0ro 0CTaTKa C C-KOH- |
|
11,a |
6b!JIO |
o6ttapy.tKeHO |
CHH.lli.eHIIC |
|
lIHq>eKlI,IIOHHOH |
aKTHBH0CTII |
II |
l(H |
|
T0IIaT0I'eHH0CTII |
[Lee |
S. |
et |
al., |
|
1989]. TaKIIe 2Ke J].attHhie 6bIJIII |
|
110Jiyqettb1 |
AJIH |
BI1O [Chakrabar |
|
ti L. et al., 1989]. |
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Pa3Mep 6eJIKOB envy BI1Y.-l II B114-2 pa3JIII'IeH, 3TII pa3JIII'mH HaII6oJiee Bb1pa2KeHbI AJrn TpattcMeM6pmrnoro 6eJ1Ka. EcJIII M0JieKyJrnpttaH Macca 3TOro 6eJ1Ka y BI1Y.-l nocT0HHHa
(41000),TO BI14-2MO.lli.CT |
co,ll.ep.tKaTb |
KaK K0p0TKYIO |
(36 000), |
TaK II AJIIHHy10 (43 000) |
M0JICKYJIY |
3TOro 6eJIKa |
[Kong L. |
et 0KHaI1988]. Pa3JIHqII5J ae BCJ!lflJHHe |
M0JieKyJibl 6eJIKa o6yc |
JI0BJICHbi II0JI0.lli.CIIIIeM TepMIIHHpyIOmero K0)l.0Ha B reHe env. Hape3aHIIe 6eJIKa-11pCJ1.lllCCTBe1rn1ma gpl60 np0IICX0J].IIT B a1111apaTe fOJib)]..lli.H B BaKy0JIHX C KIICJ!b!M 3HaqeHIIeM pH, Il03T0MY J].06aBJieHHe K 3apa2KeHHblM KJICTKaM CJJa6bIX 0CH0B3HHH (ttanpHMep, XJIOPIIAa aMMOHIIH) npIIB0J].HT K 6JIOKHpo BaHHIO ttape3aHH51 II np0AYKllHH HeIIttcpeKl(HOHHOro BHpyca [Willey eaHct 0KpeI1988]. B npoBeAeHHbIX 1-rnMII 0IIbITax A06aBJieHIIe XJI0pII,ll.a aMM0HIIH K KJieTKaM Il0lJCK o6e3b51Hbl, IIH qml(IIpOBaHHblM peKoM6HHaHTHbIM BIIpycoM 0CII0BaKl(IIHbl C BCTpoeHHblM reH0M env,TaK.tKe npHB0)].HJI0 K o6pa30BaHIIIO HeHape3aHH0ro gpl60, X0TH ero TpattcnopT K KJieToqHOH MeM6patte He ttapylllaeTCH [B3opoB A. H., ByKpIIHCKaH A. ..,A 1990]. Hape3aHIIe oeymecTBJrneT KJieT0lJHaH npoTea3a, K0T0paH II0Ka He HJ].eHTHcjJHl(HpoBaHa .YqaCTOK ttape3aHII51 npeA-
aHep
CT3BJieH BblCOKOKOHcepnanrnHOH TIOCJIC,/1,0BaTeJihHOCTbIO, o6l1LCH y BCCX peTpon11pycoB H 06Hapy1KeHHOH TaKlKe B y1IaCTKax ua pe3aHirn. fJIHKOIIpOTCHAOB opTOMHKCOBHpycoB H 11apaMHKCOBH pyccrn, Jhrn (Apr - X- Jhrn)Apr -Apr, r,11,e X mo6of1 aMHHOKHCJIOTHbli'! OCT/ITOK, a 11epT011Kll o603Ha•JalOT aJJbTCp
HaTHBHb!H aMJ1HOKHCJIOT!lhI!'i OCTaTOK. CJie,uoaaTeJibHO, 3a Ha pe3aHlle )],()Jl)KHbl 61,1Tb OTBCTCTBCHHbl pOJJ.CTBCHHb!e KJieToq Hbie npoTea3bl. B y•JaCTKC npoTeOJIH3a HaXO)].HTCH apnrn,rno
Bb!H ocTaTOK y C-KOHI\a gpl20 B IIOJIO)Kemrn 511. |
MyTaHTbI |
c 3aMeIIeHHbIM aprn1111HOBbIM ocTaTKOM [Reitz M. et |
|
1989] |
II ,n;pyrHMH OCTaTK3Mll B o6JJaCTH Hape3aHHH [Pal |
R. |
et al., |
1988] HaKaIIJJHBaIOTCH s HOpMaJihiIOM KOJllI1IeCTBe, O,LI,HaKo OHH |
l!C o6Jia)],aIOT IIHqJeKUHOHHOH aKTHBHOCTb!O B CB513H |
C |
OTCYT |
CTBHCM Hape3aHHH gpl60 H He BW3b!BaIOT o6pa30BaHHe CHH
IJ,H'rHeB [Freed E. et al., 1989]. BH4-2 co,'1,eplKHT B yqacTKe 11ape3aHH51 napy Tptt Apr, a He napy ocHOBHbIX aMHHOKHC JIOTHbIX OCTaTKOB; B03MO)Kll0, B nape3aHHe I'JIHKOflpOTCH)l,OB B}fl-I-1 11 BH4-2 BOBJieqeHhI pa3111,1e KJICTOlJHbie 11p0Tea3u
[Freed E. et al., 1989].
IlpoueccHHr 6CJIKOB env CKJI3AbIBaeTC5l H3 CJICAYIOUUIX no
CJie,IJ.OBaTeJJbHblX ::nanon: a) CHHTC3 TIOJIHIICIITHAa, BerpaHBa-
1me B 3IIAOI1Jl1:l3MaTH'ICCK!Ill penIKyJJYM, KOTpaHCJIHILHOHHOC
.uo6anJieHIIC N-CB5l3aHHhlX Bb!COKOMaHH03HblX yrJICBO)I.JlblX I\CTTOlJeK; 6) uape3aHHe npe)l,llieCTBeHHHKa gp160 Ha )l,Be cy6n
CAHHHUbl gpl20 H gp41 B cpeAHHX OT,IJ.e,1ax KOMIIJieKca fO.lJbA lKtt; B) J(HJ!bHei:iwa5l MOAHqJiiKaIJ,H5l cy61>e)l,HHHQ, 1rnnpaBJieH mrn Ha ynop5lAOlJCHUe yrneBO)l,HblX uenotJCK (y;1_aJICHHC OCTHT
KOB l'JIIOK03bl fJIIIK03H,Lla30H I H II, 4 OCTaTKOB MHHH03bl
MaHH03H,lla3ofi I H 2 ocTaTIWB MaHH03H)].a3oii II, ,11,06anJ1em1e N-aI\eTHJirJJIOK03aMI!Ha, raJJaKT03bl, CHaJIOBOH KHCJIOThl C no MOULhlO c11eu11qrn•1eCKHX rJIHK03HJITpaHC(j:lepa3 B KOMIIJieKce
ro,ilb,ll,1Kll); |
r) |
TpaHCIIOpT |
Ha KJICT01rny10 |
IlOBepxHOCTb; |
,'1,) |
BCTpa11nam1e |
B JIHTIH)].HbIH 611c,1M1 |
c |
HapylKHOH cTopoHbI |
11JJa3ManP1ecKoii MeM6paHhI |
[K.ozarsky |
K. |
et al., |
1989; |
Pal R. |
et |
al., 1989; |
Dewar R. et |
al., 1989]. |
4acTb MoJ1eKyJ1 |
gpl60 |
ycKOJib3aeT OT Hape3amrn II IJOABepraeTcH MOAH<p11uupona HH10 no aJI1>TepHaTHBIIOMY nyrn, B03Mo1K110, ll /lpyrrrx CTPYK· Typax annapaTa [OJ!b,ll,)KH. TTpH 3TOM K yrJJCBO,Ll,Hb!M 1\CIIO'IKaM A06aBJifllOTC5l tjJyK03a H raJiaKT03a, HO HC CIIaJJOBaH KHCJIOTa [Dewar R. et al., I989]. MoHOBaJICHTHblH HOHO<pop MOHeH
CHH, 11HrH6Hpy10m11n BHYTPHKJICTQl!HbJH TpaHCIIOf)T 6eJIKOB tJepe3 KOMTIJJeKC roa1b)l,1KH, B BbICOKHX KOHUeHTpau11qx (25 MKMOJJb) no.uaBJI5leT Hape3aHHC gpl60, a B HH3KHX KOH
IJ,CHTpaQH5lX ( 1-10 MKMOJ!b) 6JJOKHpyeT BTOpll'HIOe rJIHK03H JlIIj)OBaHIIC H yMeHblliclCT HHCpeKlVIOllHYIO aKTHBHOCTb. Yr,ne-
BOAHblll K0MII0HeHT Ba)KeH AJIH CTa6HJU!3aIJ)U! 11 KOppeKTH0H EOHCpop1v1a11,1n1 M0JieKYJibI 6eJIKa [Pal R. ct al., 1989].
Ha KJICTOlJHOH H0BCpXH0CTH H B C0CTaBe BHpHoHa 6eJIKH gp120 H gp41 0CT3IOTCH CBH3aHHhlMH T0J!bK0 HeKOBaJICHTHOH CBH3b!0, 3THM o6nHCH5ICTCH JICrKoe QTjJaMbIBamre gp120 OT IT0BepxH0CTH KJICTKl1 !-!JU! BHpH0Ha o6CTOHTCJlbCTBO, HMCIO· mee orp0MH0e 311aqeHHe ,Il.JIH rraToreHe3a HHq:>CKIJ,Hll H HMMYH· uoro 0TBCTa. fIOCJie YJI.3JICHHH gpl20 BHpH0Hbl H rwa3MaTH11e CKaH MeM6paHa aapa)KCHHOH KJICTKII o6oraltleIH,l gp41, Il03TO· MY aHTHTeJia np0TIIB gp41 00.il('C 3(jHjlCKTHBHO JJCHCTBYIOT B 33BHCHMblX OT aHTHTCJ[ I IIT0T0KCll'ICCl{l(X pcaKIJ,HHX II gp41 HBJIHCTCH J1Y'llllefi MHllICllhl0 )(JIH ll)!TOTOl(CHlJCCIm X T-1,JICTOK, lJCM gp120.
PerymnopHbie 6eJIKR. K HHM 0TH0CHTCH np0;'J.YKTbI pcry JIHT0pHbIX reH0B Tat, Rev, Ncf, Vif H Vpu. .3rn 6eJIKI1 3KC· npeccHpyIOTCH Ha paHHCH CTaJUIH BHpyCHOH HHq:>CKUHH, AO CHH· TC3a CTpyKTypHblX 6eJIKOB, Ii pcrym1pyIOT 3Kcnpeccmo CTpyK rypHb!X H HecTpyKrypHbIX reHoB B114. Han6o.nee «M0l!JHbIM» 6eJIKOM·TpancaKnlBaTOp0M, yaeJIH1ll!BaIOUJHM B 50-1000 pa3 TpaHcKpmrumo BHpycHoro re1wMa, HBJJHCTCR 6eJIOK Tat. Ero M0JieKyJrn coJI,ep)KHT 86 ocTaTK0B II HMeeT MoJieKynHpHyro Mac ey 15 500. Bnpyc1,1, l!MCK)!JlllC ;7,CJICIJ,HII B reHe tat, He CIIOCO0· lib! peTIJllIUMrOililThCH II BWlhIBaTb IJ,IITOH3TIJllCCKHH 3qJQ)eKT. MexaHH3M JLCflCTB!UI Tat CJl02KCH II JLO K0HIJ,a HC Bb!HCHCH.
5eJIOK |
.n.eikrnyeT na 11,11c-aKTIIBHYJO o6JiaCTb |
Tar (trans-ac |
tivating responce), pacnoJI0)KCHHYIO B LTR OT |
+ 1-ro |
JL0 rrpH |
MepHo |
+56-ro HYKJIC0THJJa. .3Ta o6JiaCTb C0Brra.n.aeT |
c 5'-He |
Tpa11c.1npyeMhlMH o6JiaCTHMII BHpycHbix MPliK c Bb1pa2KeHHof1 |
BTOpH4HOii crpyKTypofr |
[Feng S., Holland |
1988; Ding |
wall C. et al., 1989]. IIpe,IJ.mmarmor, 'ITO Tat .11.eilcTByer Ha CTaflllll IIHHILHaIJ,Hll TpaHCKlHlnIUlH [Gentz A. ct al., 1989], 3Jl0HraIJ,HrI TpaHCKp!Ull]HH, Ha nocnpaHCKpHIIUHOHHOll CTaJI,Hll HJJH Ha 06e11x CTaJI.HHX, cramrn TpaHCKpHl!IJ,l!H II IIOC'ITP3HC KpHIHJHOHHOH cTa.11.HH. Eme 0JJHcl Bepcn,1 (no-BHJIHMOMY, Ha H60J1ec JI.OCTOBepHaH), 0CH0BaHIIaH Ha pe3yJibTaTaX 0llbIT0B no BBC/lCHHI0 Tat B HApa H IUIT0I1Jla3MY 00il!IT0B, 06bHCIIHeT Mouu11,1/'i cTHMYJIHpymm11ii 3cj)q1eKT Tat TeM, 4T0 MPHK B ero rrpHCYTCTBllH XHMII'ICCKH MOJI.J!(jrnu11pyeTCH HJIH CBH3bIBaeTCH C KJICT0'!Hh!Mli QJaKTOJ)3Mll, a M02KCT 6b!Th, 11 C caMHM Tat B H.11.pe 3apa)KCHHb!X KJieT0K. TaKaH MOJI,ll(}HIUHPOBaHHaH HJil1 CBH3aH11a,1 PHK 6oJice 3QH1)CKTHBHO Tpa11c11oprnpyercH B lJH·
Ton.1aaMy II 3Kcrrpeccnpyerrn |
B pn6ocoMax |
[Braddock M. |
et al., |
1989]. |
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cocTane 6eJIKa |
npoJI,yKTa |
3K30Ha I |
BhIJ1eJIHIOTC5l |
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rrp1rnera1oma,1 K N-KOIIIW, |
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6oraTa 11,llCTCIIH0BblMH 0CT3TK3Mll. 3Ta 06.nacTb BaJirna }I.ml KoHtjiopMau:1mttttoi'1 YKJiaJJ.KH M0JJCKYJibI 6eJ1Ka H 1333HMOJJ.eiicT-
13HH c PHK-MnllleHbIO, a B03M02KH0, II ;:vrn sallIJlTbI OT KJie T011HbIX npoTea3 [Bachmair A., Varshavsky A., CMbMla1O6-
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co,n_ep.lKIIT II0CJie,ll_0l33TC.'1bH0CTH, 0TBCTCTBeHHblC 33 |
0CIIOBHYIO |
<j)yHKI IUO 6eJ1Ka 3KTHB3UHI0 .9Kcrrpecc1rn reHOB. |
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I II o6ycJIOBJIHB3CT TpaI-ICITOpTB 51)],po H CBH3blB3HHC |
c PIIK-MHWCHbIO n TaK,Ke yLJaCTByer n aKT1rnau:11n Tpattc
Kp1mu1m renon. O6JI3CTb IV yrnmmaeT 3KTHBa1uuo [Green M. et al., oMbM3B1PHJI. MYT3HT0B 6eJIK3 Tat CIIJlbH0 1rnrn611pyeT
3KTHB3UHIO nyTeM K0HKypeHUlfH C H3THBHblM 6CJIKOM. Bb!cKa3bIBaeTCH npe,nJIO/KCIIHe 06 HCl10Jlb30B3H1HI KOpOTKHX nenTH· Jl.0B, C00TBeTCTBYIO!J..(HX 3KTHBHblM 06J13CTHM Tat, B Ka11eCTBe 3HTHBHpycHblX rrpenapaT0B, HarrpHMep Jl.JlH 0JIOK!IpOB3HHH CBH3h!B311HH 6e.rrna C MlflllCHhIO H ero TpcrncnopTa B HJl.p0. CJia6bIC 0CHOB3HHH, T3KHe K3K XJIOpOKBHH, pe3K0 CTHMy.rmpyIOT ero
Jl.CHCTBHe, aauv1ma51 6eJioK Tat OT npoTeoJ1IIT1111ecKo:it JJ.erpa JI.au,mi nptt 3HJ(01UiT03e [Franl,;el A., Pabo C., CMbbCK1
BeJioK Rev c M0J1eKyJ15!ptto?i Maccoi-i OM1III1 5IB,1HCTC.H cpoc
cponpoTeH,r(oM. OH ,1J0K3.'lll30BaH B ,1,n_pe, B tJ3CTH0CTH B HJl.· pb!IUKaX sapa.lKCIIl!b!X KJieT0K 1[Falber B. et aI., 3MbMHK13KC·
npecc,rn Rev npnB0Jl.HT K sKcnpecrnn MPHK CTPYKTYPHbIX 6emrnB n CHHTKemIIo :.mcnpeccirn MPHK pery.rrnTopHbIX 6eJI
K0B, BKJII011aH ero co6cTBCHHYIO npoJ1.yKilIUO. l3eJIOK Rev He peryJrnpyeT cn.c 1aAc11Hr PHK, a Jllllllh aKTHBHpyeT sKcnopT H3 H,1pa 11YJ1a He lIOJIHOCThIO CIIJTaiicHj)OB3HllhIX IUIH Hecn.riaii CH))0BaHHh!X BnpycH1>1x MPHK, K0T0pb1e B 00I,J1m1,1x ycJIOB1rnx B u11rnnJia3MY He Tpattcnoprnpy1orrn. TeaBcj)yHKIVIII Rev Tpe-
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PHI<-nocJ1eJ1.0BaTeJI1>HOCTb |
RRE (Rev |
responce ele |
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C Bhipa)KeHHOii BTOp!PIHOH |
CTJ)YKTypoii, |
HaX0Jl.HlU3HCH |
B 06 1acTH retta env eaKH[Arrigo BTHet al., |
CMbM61Cullen B., |
Greene |
W., HMbMCH1OcTaeTC5! 11enoHHTm1M, |
B3aHMOJle:itcrny |
eT Jill 6e.r1m, Rev nenocpe,n_cTBCHII0 c 9Toii PHK 11J111 )Ke H3Me
rrneT 3KTI!BH0CTb KJICT0•IHblX 6eJIKOB, B33HMO)].eHCTBYIO!J..(IIX C lUlllirnii 11001eJ(oBaTeJ1bH0CThI0 [Cullen 1pT1Greene H0e1CMbMy
C noM0!J..(bIO MyTau110HHoro anaJittsa B cocTaBe 6eJIKa Rev
0hlJJH o6Hapy.}KeHbl ABe O0JJaCTH. ilepBaH C0JJ.epJKHT octaTKli 0CHOBHbIX aMHHOKHCJIOT, cyw.ecTBeHHbie JI.JIH TpaHcnopTa 6eJJ Ka B HJI.P0 [Perkins A. et al., 1989; MaJim M. et al., 1989]. BTopaH o6JJaCTb ocyw.ecTBJJHeT 0CHOBHYI0 TpaHCaKTHBHpy10- w.y10 cpyHKUHIO. MyTaUHII B 3THX 06.1acn1x npHBOJI.5IT K K0H Kypel!TJ-IOMYTIOJl.aBJ1eHHJO q1yHKUHH 6eJJKa JJH6o 3a CtJeT 6JJO KHpoBaHHH ero rrpoHHKHOBeHHH B 5IJI.PO H pa3o6u eHH5l TpaHC nopTBO{I cpyHKUHH OT cpyHKUHH aKTJrnau,HH, JJH6o 3a Cl!eT 6JJOKHposamrn ero aKTIIBHpy1ow.eA q:>yHKUJrn [Malim M. et al., 1988]. CTpe,11.JIOJKeHa f!IIIOT€3a O B03MO.}i{HOCTH npttMeHeHH>l MyTaHTHbIX 6eJ1KOB JJ.MI Jie11emrn 6oJJhHbIX CCTH,U.
B OTJJHtIHe OT 6eJIKOB Tat 11 Rev 6e.r10K Nef He Tpe6yeTcH ,ll)IH penJIHK8UHH B119 in vitro. 3TOT 6eJIOK C MOJJeKYJJ5IpHOI{ Maccoi1 27 000 MHpHCTHJIHpoBaH H CBH3aH C l.I,HT0fJJJa3M3TH t{CCKHMH MeM6paHaMH K.JJeTKH. OH TIOJI.3B.15IeT TpaHCKpHilll,HIO
m1pycHblX reHOB, peryJittpyeMyJO LTR [Niederman M., |
1989], |
11 q:>yHKUHOHHpyeT KaK penpeccop rncnpeccHH reH0B |
BH9. |
TaKHM o6pa30M, 3T 6eJJOK H3XO,II.HTC5l B KOHKypeHTHblX B33HMOJI.eiicTB1rnx c 6eJ1KOM Tat. Bo3MO:iKHO, Ncf cnoco6cTByeT ycT8HOBJJeHHJO .JJaTCHTH0fO COCTOHHHH HH<peKJ.]JIIl.O,u.HaKo, HH rn6Hpy10w.ee JJ,e{JCTBHC Ncf na TpaHCKpl!IIUl!JO Bblp3)KeH0 He MHOrO cJJa6ee, lJeM CTHMyJrnpy1ow.u11 scpcpeKT Tat: Nef crroco6eH D0)UHlJIHTb TIH111CK})IIIIUIIJ0 JIUlllb B 2-4 pa3a [Cullen B., Greene W., 1989]. l'i1roJ1cir11 1 recKaH pOJJb 3T0ro 6eJ1Ka B lIH qJeKUHOHHOM IIJIKJic B114 ocTaeTCH Hcs:i:cHoit. BeJIKH Vif H Vpu II3YtJeHbl M3JI0. OHH Hrpa!0T pOJlb B MOpcporeJ-Je3e II C03peaa HHH BHpycHOH tJaCTHU.bl. BeJJoK Vpu HBJIHeTeH cpocqionpoTeH ,11.oM c MOJJeKYJIHpHoii Maccofi 16 000. OH CHHTe3upyeTCH Ha pH60COMaX, CBH3aHHblX C MeM6paH3Mll, BCTp3HB3eTCSl B MCM6paHbl 3H.LJ.OnJia3Man1 11ecKoro peTnKyJiyMa. Ero cpyHKUHH Ba}KH3 .Ll.JI5! c6opKI1 JI.HH 1101IK0133HHH B11pyc1rnii '13CTHUbl. Bo3MO)KHO, OH JI.eikTByeT no HenpHMOMY MexaJ-JH3MY, HHJI.YUHPYH ('llHTe3 KJ1€TOtJH0ro cpaKTopa, cymecTBCHHoro )].JIH ITO<IK0BaHHH [Strebel K. et al., 1989]. Ilo CBOeH cpyHKUHH H JioKaJJtt3aUHH B MeM6pauax 6eJIOK CXO.Ll.CH c MaTpHKCHblMH 6e.JJKaMH mrpycoa rpnnna 11 napaMHKpoBupycoa H c 6eJ1KOM NS28 pOTaBHpycoa (cM. rJJaay 7). OTCYTCTBlfe 3T0f0 6eJIK3 H.JJH ero JI.ecpeKT MO )l(CT T0pM03HTb Il0tJK0B3HHe H rrpOJI.YKUHJO BHPYCHblX 'l3CTHU TaKUMH KJieTKaMH, KaK MaKpocj:JarH H MOH0UHTbl. HaKOHeu,
Vpr HBJIHCTCH 6eJIKOM c HeH3BeCTHOH cj:JyHKUHeii (Ta6JJ. 19). Mop(f>oreHe3, C6opKa BH9, KaK 11 JJ.pyrnx o6oJiocieqHbIX
BHpycoB, npouCX0,11.HT Ha IlJl33MaTHtieCKOH MeM6paHe KJieTKH, KyJI.a He33BHCHMO ,npyr OT ,upyra TpaHcnoprnpyJOTC5l Bee K0M HOHCHTbl nttpycHoi'.iqacTHI.fbI. BeJJKH env cnHTe3npyIOTCH Ha CBH3aHHbIX c MeM6pattaMu pH6ocoMax 11 rJIHK03HJJHPYIOTCH no
