Recen t Advances in the Techn iques of Prote in-prote in In teraction s

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1 14 1 V o l114,n o Ch inese Jou rnal of B iochem istry and M o lecu lar B io logy Feb 1998 :, (, ) Recen t Advances in the Techn iques of Prote in-prote in In teraction s H uang Cu i2fen Ye Q i2n ong (B eij ing Institute of B iotechnology, B eij ing ) Abstract P ro tein2p ro tein in teraction s are im p licated in every cellu lar p rocesses such as DNA rep lication, tran scrip tion, tran slation, sp licing, secretion, cell cycle con tro l, signal tran sduction and in term ediary m etabo lism Som e in teraction s in cells last long and o thers are tran sien ṫ T he specificities of these in teraction s are essen tial fo r the function s of p ro tein ṡ P ro tein2p ro2 tein in teraction s can alter the k inetic p roperties of p ro tein s, resu lt in the fo rm ation of a new b inding site, inactivate a p ro tein, and change the specificity of a p ro tein fo r its sub strate In the past few years, very rap id advances have been m ade in iden tification and characterization of p ro tein2p ro tein in teraction ṡ H ere, bo th classical and, especially, recen t m ethods to study p ro tein2p ro tein in teraction s w ere summ arized F irst, b iochem ical m ethods such as p ro tein affin ity ch rom atography, affin ity b lo tting, imm unop recip itation, cro ss2link ing w ere review ed; Second,mo lecu lar b io logical m ethods such as p ro tein p rob ing, phage disp lay, tw o2hyb rid sys2 tem w ere described; F inally, a new b io sen so r techno logy based on su rface p lasm on resonance, w h ich have becom e increasingly popular fo r the qualitative and quan titative characterization of p ro tein2p ro tein in teraction s,w ere in troduced Generally speak ing, the b iochem ical m ethods requ ire the purification of p ro tein s and (o r) an tibody p reparation, w hereas som e m o lecu lar b io logical m ethods such as tw o2hyb rid system do no ṫ T he m o lecu lar b io logicalm ethods w ere developed based on con struction of gene lib rarieṡ T herefo re, the coding sequence that in ter2 acts w ith a know n p ro tein can be directly deduced in com parison to the b iochem icalm ethodṡ T h is review em phasizes the p rincip les, app lication s, advan tages and disadvan tages of the techn iques m en tioned above Key words: P ro tein, In teraction, T echn ique [ 1 ] : , : China Academic Journal Electronic Publishing House All rights reserved

2 ; ;, (p ro tein affin ity ch rom atography) (affin ity b lo tting ) (imm unop recip itation) (cro ss2link ing), (p ro tein p rob ing) (phage disp lay) (tw o2hyb rid system ) (su rface p lasm on resonance), 1 : Sepharo se,,, (SD S) :M ason [ 2 N u sb S10 ] S10 A ffi2gel,,,, SD S2PA GE N u sb W estern b lo tting, N u sb S10 : (1) 10-5 m o lgl ; (2) ; (3) ; (4) 2 W estern b lo tting [ 3 ] SD S,, ( ) 3, :,,, SD S2PA GE W estern b lo tting : F ilho l g p 53 [ 4 ] g p 53 sf9, 3,,, p 53 A (SPA ), SD S2PA GE W estern b lo tting p 53 : (1), China Academic Journal Electronic Publishing House All rights reserved

3 ; (2), ; (3) ; (4),,, 4 ( R SSR, R R,, R SH, R SH ), (D T T ), : [ 5 Gunzbu rg R as 60kD ] 35 S, D SP (dith io2b is2 succin im idyl p rop ionate),,,, R as, SPA D T T, SD S2PA GE, R as 60kD 5 :, : B lackw ood [ 6 M yc M ax ] M yc 125 I,, M ax : (1) ; (2) ; (3), 6 (F ig 1) : g g, [ 7 ], (96 ),, [ 8, 9 ], : (1) ; (2) ( ) ; (3), : (1) DNA China Academic Journal Electronic Publishing House All rights reserved

4 ; (2), ; (3) ; (4) D (DD ) DD DD DD, Balbgc, m RNA, cdna, cdna DD, F ig 1 Schem atic m ap of phage disp lay DD Phage2EL ISA, DD EL ISA 10 7 tfugm l EL ISA tfugm l, DD DNA, cdna 729 bp, V H 354 bp, 118 ; VL 327 bp, 108 ; V H VL (Gly4Ser) (in teraction trap ) [ 10, 11 ] (F ig 2) :, DNA (A ) (X ), A + X DNA (B ) (Y), B + Y 2, L eu lacz DNA, DNA (up stream activating sequence, UA S) lexa, DNA L eu L acz, X Y DNA UA S lexa L eu, X2gal, B + Y,, Y Y X China Academic Journal Electronic Publishing House All rights reserved

5 Jun Fo s TN FR FAN So s [ Grb s ] ; ; ;, :, :, 1 Λm o lgl ; : F ig 2 Schem atic m ap of two2hybrid system ( a) If X and Y p ro teins do no t interact, transcrip tion of repo rter genes w ill no t be activated (b) If X and Y p ro teins interact, tran2 (1) scrip tion of repo rter genes w ill be activated, A : DNA binding dom ain; X: interest p ro teins: B: DNA 2transcrip2 tion activation dom ain; Y: screening p ro teins ; (2), X Y, ; (3), ; (4) (, ) (5), CPP32 15 CPP32 ( yam a, apopain) ICE, (death p ro tease), (death sub strate) R T 2PCR CPP32, DNA, CPP32 pgbt 9g CPP, cdna DNA pgbt 9gCPP cdna CPP32, Β2 42 Β2, 5 1 h 5, E coli HB 101,, cdna CPP32 (15 ) China Academic Journal Electronic Publishing House All rights reserved

6 CPP32 8 [ 17 ] : ( ) ( ),,, : T T (TCR ) (M HC) (an tigen) M argu lies [ 18 M HC an tigen TCR M HCgA n tigen ], M HC A n tigen, TCR M HCgA n tigen T : (1), 1 10 Λg 50 ; (2) 10 m in ; (3) ; (4) ; (5) : (1), ; (2) 9,, [ ] ; References 1 Ph izicky E M, F ields S P ro tein2p ro tein interactions: m ethods fo r detection and analysis M icrobiol R ev, 1995, 59: M ason S W, L i J, Greenblatt J D irect interactions betw een two E scherich ia coli transcrip tion antiterm ination facto rs, N usb and ribo som al p ro tein S10 J M ol B iol, 1992, 223: Sam brook J, F ritsch E F,M aniatis T M olecu lar C loning, A L aboratory M anual, 2nd ed, N ew Yo rk: CSH, F ilho l O, Baudier J,Delph in C, L one2m ackenbach P, Cham baz E M, Cochet C Casein k inasee g and the tumo r supp res2 so r p ro tein p53 associate in a mo lecular comp lex that is negtively regulated upon p53 pho spho rylation J B iol Chem, 1992, 267: Gunzburg J D, R ieh l R,W einberg R A, Identification of a p ro tein associated w ith p21 ras by chem ical cro sslink ing P roc N at A cad S ci U SA, 1989, 86: B lackwood E M, E isenm an M A helix2loop2helix zipper p ro tein that fo rm s a sequence2specific DNA 2binding comp lex China Academic Journal Electronic Publishing House All rights reserved

7 w ith m yc S cience, 1991, 251: Co rtese R, Felici F, Galfre G, L uzzago A, M onaci P, N ico sia A Ep itope discovery using pep tide libraries disp layed on phage T rend s B iotechnol, 1994, 12: Jespers L S, Roberts A,M ah ler S M,W inter G, Hoogenboom H R, Guiding the selection of hum an antibodies from phage disp lay reperto ires to a single ep itope of an antigen B iogt echnology, 1994, 12: W inter J Bacteriophage disp lay: pep tide libraries and drug discovery D rug D evelop R es, 1994, 33: F ields S, Sternglanz, T he two2hybrid system: A n assay fo r p ro tein2p ro tein interactions T rend s Genet, 1994, 10: A llen J B,W alberg M W, Edw ardsm C, E lledge S J F inding p ro spective partners in the library: the two2hybrid system and phage disp lay find a m atch T rend s B iochem S ci, 1995, 20: Chevray P M, N athans D P ro tein interaction cloning in yeast: identification of m amm alian p ro teins that react w ith the leucine zipper of Jun P roc N atl A cad S ci U SA, 1992, 89: A dam 2k lages S, A dam D,W iegm ann K, Struve S, Ko lanusw, Schneider2M ergener J, Kronke M Fan, a novelwd 2repeat p ro tein, coup les the p55 TN F2recep to r to neutral sph ingom yelinase, Cell, 1996, 86: Chardin P, Camonis J H, Gale N W, A elst L V, Sch lessinger J, W igler M H, Bar2Sagi D H um an So sl: a guanine nu2 cleo tide exchange facto r fo r Ras that binds to GRB2 S cience, 1993, 260: Ko lanus W, N agelw, Sch iller B, Zeitlm ann L, Godar S, Stock inger H, Seed B ΑL Β2 intergringl FA 21 binding to ICAM 2 1 induced by cytohesin21, a cytop lasm ic regulato ry mo lecule Cell, 1996, 86: Chou J, Ro izm an B H erpes simp lex virusl r1345 gene function, w h ich block s the ho st response to infection, m ap s in the homo logous dom ain of the genes exp ressed during grow th arrest and DNA dam age P roc N atl A cad S ci U SA, 1994, 91: M alm qvist M B io specific interaction analysis using bio senso r techno logy N atu re, 1993, 361: M argulies D H, P lak sin D, Kh ilko S N, Jelonek M T Studying interactions invo lving the T 2cell antigen recep to r by sur2 face p lasmon resonance Cu r Op inion Imm unol, 1996, 8: Stanger B Z, L eder P, L ee T H, Kim E, Seed B R IP: A novel p ro tein containing a death dom ain that interacts w ith Fasg A po21 (CD 95) in yeast and causes cell death, Cell, 1995, 81: M atsum ine A, O gai A, Senda T, O kum ura N, Satoh K, Baeg G H, Kaw ahara T, Kopbayash i S, O kada M, Toyo sh im a K, A k iyam a T B inding of A PC to the hum an homo logue of the D ro soph ila D iscs large tumo r supp resso r p ro tein S cience, 1996, 272: Staib C, Pesch J, Gerw ig R, Gerber J, B rehm U, Stangl A, Grumm t F p53 inh ibits JC virus DNA rep lication in v ivo and interacts w ith JC virus large T antigen V irology, 1996, 219: Sh ivakum ar C V, D as G C Interaction of hum an po lyom avirus BK w ith the tumo r2supp resso r p ro tein p53 O ncog ene, 1996, 13: China Academic Journal Electronic Publishing House All rights reserved