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Homologies in procaryotic ribosomal proteins: Alanine rich acidic proteins associated with polypeptide translocation
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HOMOLOGIES IN P R O C A R Y O T I C RIBOSOMAL PROTEINS: ALANINE RICH ACIDI:C PROTEINS, .ASSOCIATED. WITH POLYPEPTIDE TRANSLOCATION ..... Louis Y. V I S E N T I N , Alas~air T, MATL-tDEOl~ m ~ d M a k o t o - Y A G U C H I ..
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R-ece~vefl 2B 3anx~ary ] 974
1. I n t r o d u c t i o n
.~ " s t e a r o t k e r m o p h f f u a ~tr,Mn ] 0 were prepared as dez] cfibe.d pirevi0aasly 1.17] a n d the. acidic proteins s.e]eclively :extracted by. t h e m e t h o d s ,of H a m d , Ko]Ca and Trmasloeafion oft'he growing 1~pt~,de chain ,~n.ihe Na]camol,o- [ J_3}. The three pzot,e~r~s liberate d by this sxcnthm;is o f proteins is m e d i a t e d t o ~ome e x t e m b y p r o c e d u r e were ,co~aeentrated l~y px.eeip]'ta~ion wi,*,da certain n o v d acidic :ribosolnal pr.o~.~eins [J~-I 6 ] . These 2.25 v:ol. o f - - 2 0 C acetone a n d .1.he L T - - L 1 2 e.quivaproteins, :designated L7 .and L 1 2 i.n E s c h e r i e h b 2 ,¢oli, lents separated and purified ~s described f o r / ~ e o l i b y •are loeMized on the 5 0 S ,ribosomal ;subunit and :differ only in the acetylation state , o f t he .N-,term~nal serJne. Mbll:er ,et al. l a ] . Their p u r i t y was established b y a 2[2]. T h e enistence ofstruc,tuxal h o m o l o g u e s t o ihese dimensional zcrylamide e]e.ctrophoresis_ m e t h o d Z~nialanine rich, acidie proteins in Ja-t:ost proeaxyotic and ]at t:o thai descfibedb.yKaTtse~hrnidt and W i t t m a n n [ 18] eucaryotic cells has b e e n .sugge.sted b y r e c e n t haami~no~xe.ept that the first dimension di~.cs, ~ n at p H &6, logical expefimen,ts b y 1. G. W,oo] and G, Stbffler (perwere 8% w i t h r e s p e c t . t o ae~3z]arMd~_ T i ~ h o m o l o g y sonal c o m m u n i c a t i o n ) . a n d ~ d e n t i l y o£~the L 1 2 - - L 7 equivalents f r o m 2~. Their ~abiquity as ri,'bosomal co~.stitu.ents indicate a stear, a t ~ h e r m o p h i l u s wras eolafirmed b y L~e irrmaunohigh degree of.~onse:rvation in these mo]eetfies during cheJ~fical reactivity o f t h e is.olated pm'e prot:eh~s t 0 the evolution ,of the ribosome. ~le~ce, ,comparative ,lhe anfisera p r o d u c e d in N e w Z e a l a n d red rabbits structural and functional analyzes o f these unusua~ against purified E. eol~ L J 2 115]. This w a s ~ u r t h e r po!ypepfldes c o u l d reflect b o t h the; gmaetie modifi.evidenced b y am,hao acid. analysis in a Daaxnam Z)-5'O0 cations possible in the .evolution o f 'the r i b o s o m a l .~m'* a m i n o acid analy-sex~ fcallowSng pr.o~tein ~ y d r o l y s i s fo,x :as well as yield ~ome ~nsight in*,o Che n a t u r e ,of-the .'ac2 4 and 7 2 ~ J:n ,6 N H C I a t 11,O~:C, as well ,as b y tive sites" wi,t~n the molecules. As a firs~t p.:o~ession sequence analysis b y a u t o m a t i c Edmma d e g r a d a i m n 1~19] fm a B e c k m a n Model ,890 :C Sequenc,er. 'For sein this direc tion 'we .report ]aerein the ~solafion, ,chemiquence f a n s the.,t~az01inone derivatives w e r e b y . cal ,characterization and N-terminal s e q u e n c e analysis ,drolyzed separately With 6 N H C 1 and H L [20] a.t o f an L 7 - - L i 2 equivalent frora 13. s ; . t e a r o t h e r m o p h E u s . . We c o m p a r e the structt., .~al features wJfla th~ ~. c o l i -. 1 3 0 ,C f o r :22 hr, mad t h e a m i n o :acids £ ~ r m e d w e r e polypeptJdes, a c o m p a r i s o n that is o f par.fieulzr n o t e -. analyze d wJfla ~'a.e a m i n o . a c i d maa!2~;'~ex-i]~J.o"lee'alax -. " in ~iew o f the consider.able in~erest.in 1he structure, : . " . weights..were detemfin ed b y t h e d o d e c y l ~Sulfate g d ~ . function a n d evolution ~of:the ge~e:t~c imn~]ation. ' • -_ el=ectrophorefic proe:edure, of;~Veber:and.,Osborn I21] o. apparatus. -.- . . . . " .:. : . -- . . . "
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o n e •batch. o f ~:clts~to ~ n 0 , h e l d e p e n d i n g appar,en'dy o n -"the g r o w t h cOnditiOns, Similar ~¢ariafionshave. b e e n r e p o x t e d f o r E . coli t,7 a n d •L12-12:0]: s i m i l a r ~o f a e m o b i l ~ W , o f E. coil ~;I--L12, o n ~w,o ,dir~er~i.on~ .eleetr.o~horesis p r o t e i n s A~ ia n d A~ f r o m B. _s,tearothermoFhi/u~ m i g r a t e t o a positi,on z h o w n in fig. 3. -A~-a. moaeeu]ar lev61,, flg.e,z c e t y l a f i o n s~ate o f 'the N - t e r m i n a l sen~ae xv/;, c o b L 7 a n d L1 2 ha~ b e e n ~ugges~ed .as~ s e n t i a l fo:r t h e p r o p e r / u n e t i o n i n g o f Ihese pro~eL~s k~ EUG a n d E F T d e p e n d e n t GTPase .~e~cd,onz [ ' 3 , 4 , 1:0i]~ W h e n ~he N - ~ . e ~ i n a l region .of fhe p u r i f i e d A~ pr,o~eins f ; o m B . ~t~m~othermophi~us was s.eq~eneed the N 4 e r m ~ a l a,e~idue was f o u n d t o be. rne~hi,onine za'ther than ~e~me. T h e f i r s l 1-S r e s i d u e s oi fl~e.A2 p r o t e i n o r B . s~ea.~ozl~ez~ophit~s me: Ne~-Thr- Lyz-'G]u-G~]n-Ile-1]e-Gln-Ala-N'N- L ~ - A s n - ~.e,l.... " - • .... . . Se~-?%~aL. . "Fig.2. Immanochemicai ~ , ~ i ~ :of/ft. ~t,earo~h:~moFh$~u~ ~ ' i e absen¢e o f a n . N - ~ e r m h a a l ~er~me mad the prepr,ot,eins t h a i , c o z r e ~ p . 0 n d 1.0 E. ,egli t~r0;teins L7,~L12. ,tempe " " s e n c e o f ~ : m 6 ~ h i o n i n e , a~ first ~uxpxisixag, it n o t u n l i k e we]l eonlnaned.An~ E. ,coli L12..~2) B..stearo~]J-.ezmOpBffus " th~ .da~-a r e p o t t e d f o r ithe ~ ¢ o h o l ~ e h y d r o g e n a s e f r o m A~ ~ 1 mgjma an ph:o~pha~e b~ff~rvxt saline pH 7;0 t l) 9:9% ,,sai|i:ne, ~. st,earo,'.tI~erma)pN:lus [23] w h e r e ~he ace~.y]a'ted
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!r~= ~. 4. Cerspaison o f ~h.e N-t~;m~na] arn~n~ a ~ d s~q~ences in i T - - L ] 2 f~om ~. ~ 1 i and B. s¢~ro;her.~.nop~jh'.s A2 pro~e~. "Ih~ s . ~ e n c e s a:e a~gned ~o ~ha~ ~aline-] g {~,~f. ~3] ) cor~eapongs !o ~'a~ine 15 in B. ste=zotTJe~opl_,iN~ A~. R~id~aes e n e : ~ e d by d ~ b l e lines a~e idenfi~M.
s erine is also r e p ] a e e d b y a r a e l h i o n i n e . F u r t h e r m o r e ,
F. Rollin a~d }~ Tess[ez fox expert ~e~b_nScal ~ s i s ~ c e
it .is o f in~eres~ @a~ ~he a l a n ~ e A.ch a cidi~ p~o~eins frown ihe ~xt~enae h M o p ~ o b~c~e~nrn, iz]~'!,~b~ci,e1"i,1~n
and to D~. H. Jennings m~d M~s. A. NaltSn fox help w~th the immunological experLn~.en~s. We :~ank D n
~ t i n ~ . b r a n ~ , also has ~ rnet~hionine W-~errninal (O,aa e~ al. ~ n p n b ] i s h e d o b s e r v a t i e n s ) . F o r t h e A p~ote~ns f~.om ~ co1$ a n d ~ s~e,a r ~
1-Ia_~'ey Kaplmn f ~ ~his v~/~a~ble discu~sioDs.
the~op1~ilus, ,it ~s possible
References
l:e s h o w ]~,om,o,'].ogies m the c h a i n s :f , h e m e t h i , o n m e N - t e n n m m of the/~, s 2 . e ~ t h e ~ a o p h i I u s pr,o~eh.~ ~ a]igned w i t h ~he iso].eacin,e-2 ,the ~.. c o g L 7 o~ L 1 2 p~,o~e:ns [f~g 4 ] . I f d-Bs is
done then 1i3 o u t o f 1he f i ~ t 16 ~esidu~s :(62.5%) are iden.~eal in ~la~ ,two e h ~ n s . I n t h o s e ~h~'t ,difl?~r.Tne,t1, glu-4 and gha-tg, c o u l d h a v e arisen b y poSnt rnuta~.on and the fizs,t t w o are as w.e]l, e,0nse~ad~e ehan~es.
S ~ a ~ h,omolopAes bm~e been shown fn: some 30 S p~ot,e'ms [24], Howeve:, ~he degree t.o which this h ~ molegy ~s ,~ontinU.ed in the primary strac~u~e o f th:~ A,~ and Aa pro~eh~s awa~s fu;the~ sequence ma~lyses o f these pro[eir~s :_~ B. sIe~ro.~,ermcphflus. ]n view o f ~ e homologies m primary s~ra~m:e and N - s ~ i b s ~ t u t i o n it ~s poss~Me t h a t ,theN - t e n n i n ~ l ,~e~ o n o f ~these p~ot,eins has i m p o r t a n t £uncfi.onat ~o]:es. .Ther.efo:e in ol,fl,~ t o e]uci,fla~e t h e .~ole of ,%hese p~ly~ e p f i f l e s ~ t o ~ ~ibosom~l ~une~ion i'ti~ n e c e s s a r y •t h a t t h e c h e n d e a l _~r'ope:des o f ~the amin.o t:erminns :be d e t e r m i n e & Si~eh s~u.dies h a v e n o w b e e n iNtia:e.d ,on: laboratory.
Aeknow]edgementsT h e a,i~.tho~ are i n d e b t e d ,to :C, tSoy, W; Row~o~-ne,
~1972) :E~..]. B~ochem. 2 5 . 5 - - ] 2 . ].2] T~e:;]~z-~, C-, "WitI~rsn.-I.Se~.D]d, B. a:nd ~]i~]te;, 'W. ~1'972) ~En;. 3. BSache:~.. 2 5 . 1 3 - t 9 . I 3 ] Te!hozs~, C., MSIle~, W., laursen, R. and Ni~!rnann15ebm]d, B. {1992) iFEBS ke~e~s 2g, 3 2 5 - 3 2 g . ]4] T e : h e ~ , C., M~51]~z, W., L~ursen, R. and ?,i~rnann]_SebNd, B. f1973) E~z. J. Biochrenl. 34, 13B-~52~5] Bre~, N., ~amasa.ki, E., Redfield, B. and WeSsshach. ]]. ( t 972) .A~ch. Biuchem. Bi~phys. ] 48, ] 48-- ] 55. I5] B~m, N. an~t W ~ s b a c b , H. ~]972~ Bio~hern. Biophys.
11~ C~n~mn~. 49, 673-~79. ]7] ~v'f~hsb~ch, 1-1.,~¢.d~e]d, ]3, Yamnsaki, E., D~",i~. :R. C:., P~k~, N. and Brow, N. [1972) A~c~h.~och~m. BSophys. 149, 110- 117. 1~] K~P.g, l-].-, ,, z,0x, 3. E., $pear~, C,, B~e~, ?.,'. and Weissbaeh, H. '[1973) L B]~I. Churn. 248, 5 0 ! 2 - - 5 0 t S]N} BIo't: N., Marcel R., C~ap.~, L- and W~ssb~ch, N- {1~73) A~eh. Bier-hera. Biophys. t 55: 4 7 5 - 4 7 7 . B~o=Nern. BiophN~ Res. Cornr~¢_~-~47, ~66--~873. ~ ] ~ Chin~li,O. and ]~a~/n~-g=gian~, A. '119739~-~r.3. iBieahern. 32,463 -472. I32~ Nnrnel, E. anti Nakalno'Io,~. 11972) 3. BID3. Chef-. 247, 6810-15817o i~5] l-l~rnei, E., XoLa, ~l. n~,~/~akar~.e]p, T. ~]97~2) J. ~ o l . .Chen~. 247, guZ.--B 14. P- -(a9;~.~ Biochem. 33~ophym llem Cormn'~r~ 46, 238"24~. "
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[ i 5 ] H i # l a n d , " J..H,~ B o d t e y ~ J ~. ~#;~.; ' G o r d o n , : J ,, ~ l a s e n b a n k , ~ . . ~.[20] S r r f i t ~ O , , G i b s * n , -D~ ~F&m~ng, E , M . , G 0 o d f l i ~ h : , " a n d .$tiscfler, G . (I97~.3). :Pre-e; N ~ t f l . . A e a d . GeL U . S . 7.(i, : ' . " --: i~ R..M.~ G i l m a n , J. G . '.and B a B a m ~ n e , D., L . t 1 9 7 1 ) -~147-1$0. ~i .. " " ~:.. !.:: " • " " .- B i o d h e r r h s t r y ] 0 , 49a~-ag,2~. l~i6.1. S c h f l e r . i ' . L , M a a s s e n , ] ; . A . a n d M ~ t l e r ; W~ ( 1 9 7 3 3 ..., . ~i t21] Weber, K. andOsbom, M . ( , i 9 6 9 ) J . B i o l . C h e i n . 244~ ' . Bioclne'nxB~ophy.~ Re~ .Gommun. 53,:90--98. ..... . . --4406~.4412, |17:] C h o w , C. T..~ ~ ' i s e n t i n , L. P . , M a t h e s o n . . A . ~F; a n d _ [ 2 2 } - . . ~ ' ~/ a a . P . . K,ttri~nd. C . ;G_..Dexxgser. E - , .~a~,re~oex;J-, Yagucl~j. M . ( 1 9 7 2 3 Blochlm. Blophys. A c . a . f i g T , . • ' ~tasc~e~, ~.~, L~t~/f~'ler, G.. a~.d W i t t _ m ~ , ~]. G.(xD733 270--:28.1: - " " " • " : . - "- i . . : . . . --Na~.ure l ~ e w ~B~01. 2 4 2 , 4 7 - - 4 9 . [ 1 8 1 K a ! t s c l m ~ d : ~ , E . m ~ d w)l~m~.n.n., 14. G° ~197'~):A'na L I 2 3 ] B r i d g c n , 3 . , K o I b , ~ . a n d ~ a r f i ~ , J . I. ( 1 9 7 3 . ) : F E ] ~ S • Bi0chem:36,401-412,,. " " " i .- " ..... • I~et,t.er~BB, l - B . ..- " " [19.1 E d m a n ; P. :and BL~gg,.G. ( 1 9 6 7 ) ] { a r . J / B k , e]aem~ I , •i[24] .3?aguclfi, ~ d . , l ~ o y , C., 2VlaZhes~n~ A . T . z l a d V i s e n f i n , . g9--91~ . " L~ P. ( 1 9 7 3 ) . C a n . J - Bioclaeaz~:ti], 1 2 1 5 - - 1 2 1 7 . • .
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HOMOLOGIES IN P R O C A R Y O T I C RIBOSOMAL PROTEINS: ALANINE RICH ACIDI:C PROTEINS, .ASSOCIATED. WITH POLYPEPTIDE TRANSLOCATION ..... Louis Y. V I S E N T I N , Alas~air T, MATL-tDEOl~ m ~ d M a k o t o - Y A G U C H I ..
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D Ms;.on o f Bfolo~calSciences~ ~ a a o n a l Researeh .Counc~7 o f Canada, Ottawa, ~anocta K J A O R 6 . _
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R-ece~vefl 2B 3anx~ary ] 974
1. I n t r o d u c t i o n
.~ " s t e a r o t k e r m o p h f f u a ~tr,Mn ] 0 were prepared as dez] cfibe.d pirevi0aasly 1.17] a n d the. acidic proteins s.e]eclively :extracted by. t h e m e t h o d s ,of H a m d , Ko]Ca and Trmasloeafion oft'he growing 1~pt~,de chain ,~n.ihe Na]camol,o- [ J_3}. The three pzot,e~r~s liberate d by this sxcnthm;is o f proteins is m e d i a t e d t o ~ome e x t e m b y p r o c e d u r e were ,co~aeentrated l~y px.eeip]'ta~ion wi,*,da certain n o v d acidic :ribosolnal pr.o~.~eins [J~-I 6 ] . These 2.25 v:ol. o f - - 2 0 C acetone a n d .1.he L T - - L 1 2 e.quivaproteins, :designated L7 .and L 1 2 i.n E s c h e r i e h b 2 ,¢oli, lents separated and purified ~s described f o r / ~ e o l i b y •are loeMized on the 5 0 S ,ribosomal ;subunit and :differ only in the acetylation state , o f t he .N-,term~nal serJne. Mbll:er ,et al. l a ] . Their p u r i t y was established b y a 2[2]. T h e enistence ofstruc,tuxal h o m o l o g u e s t o ihese dimensional zcrylamide e]e.ctrophoresis_ m e t h o d Z~nialanine rich, acidie proteins in Ja-t:ost proeaxyotic and ]at t:o thai descfibedb.yKaTtse~hrnidt and W i t t m a n n [ 18] eucaryotic cells has b e e n .sugge.sted b y r e c e n t haami~no~xe.ept that the first dimension di~.cs, ~ n at p H &6, logical expefimen,ts b y 1. G. W,oo] and G, Stbffler (perwere 8% w i t h r e s p e c t . t o ae~3z]arMd~_ T i ~ h o m o l o g y sonal c o m m u n i c a t i o n ) . a n d ~ d e n t i l y o£~the L 1 2 - - L 7 equivalents f r o m 2~. Their ~abiquity as ri,'bosomal co~.stitu.ents indicate a stear, a t ~ h e r m o p h i l u s wras eolafirmed b y L~e irrmaunohigh degree of.~onse:rvation in these mo]eetfies during cheJ~fical reactivity o f t h e is.olated pm'e prot:eh~s t 0 the evolution ,of the ribosome. ~le~ce, ,comparative ,lhe anfisera p r o d u c e d in N e w Z e a l a n d red rabbits structural and functional analyzes o f these unusua~ against purified E. eol~ L J 2 115]. This w a s ~ u r t h e r po!ypepfldes c o u l d reflect b o t h the; gmaetie modifi.evidenced b y am,hao acid. analysis in a Daaxnam Z)-5'O0 cations possible in the .evolution o f 'the r i b o s o m a l .~m'* a m i n o acid analy-sex~ fcallowSng pr.o~tein ~ y d r o l y s i s fo,x :as well as yield ~ome ~nsight in*,o Che n a t u r e ,of-the .'ac2 4 and 7 2 ~ J:n ,6 N H C I a t 11,O~:C, as well ,as b y tive sites" wi,t~n the molecules. As a firs~t p.:o~ession sequence analysis b y a u t o m a t i c Edmma d e g r a d a i m n 1~19] fm a B e c k m a n Model ,890 :C Sequenc,er. 'For sein this direc tion 'we .report ]aerein the ~solafion, ,chemiquence f a n s the.,t~az01inone derivatives w e r e b y . cal ,characterization and N-terminal s e q u e n c e analysis ,drolyzed separately With 6 N H C 1 and H L [20] a.t o f an L 7 - - L i 2 equivalent frora 13. s ; . t e a r o t h e r m o p h E u s . . We c o m p a r e the structt., .~al features wJfla th~ ~. c o l i -. 1 3 0 ,C f o r :22 hr, mad t h e a m i n o :acids £ ~ r m e d w e r e polypeptJdes, a c o m p a r i s o n that is o f par.fieulzr n o t e -. analyze d wJfla ~'a.e a m i n o . a c i d maa!2~;'~ex-i]~J.o"lee'alax -. " in ~iew o f the consider.able in~erest.in 1he structure, : . " . weights..were detemfin ed b y t h e d o d e c y l ~Sulfate g d ~ . function a n d evolution ~of:the ge~e:t~c imn~]ation. ' • -_ el=ectrophorefic proe:edure, of;~Veber:and.,Osborn I21] o. apparatus. -.- . . . . " .:. : . -- . . . "
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o n e •batch. o f ~:clts~to ~ n 0 , h e l d e p e n d i n g appar,en'dy o n -"the g r o w t h cOnditiOns, Similar ~¢ariafionshave. b e e n r e p o x t e d f o r E . coli t,7 a n d •L12-12:0]: s i m i l a r ~o f a e m o b i l ~ W , o f E. coil ~;I--L12, o n ~w,o ,dir~er~i.on~ .eleetr.o~horesis p r o t e i n s A~ ia n d A~ f r o m B. _s,tearothermoFhi/u~ m i g r a t e t o a positi,on z h o w n in fig. 3. -A~-a. moaeeu]ar lev61,, flg.e,z c e t y l a f i o n s~ate o f 'the N - t e r m i n a l sen~ae xv/;, c o b L 7 a n d L1 2 ha~ b e e n ~ugges~ed .as~ s e n t i a l fo:r t h e p r o p e r / u n e t i o n i n g o f Ihese pro~eL~s k~ EUG a n d E F T d e p e n d e n t GTPase .~e~cd,onz [ ' 3 , 4 , 1:0i]~ W h e n ~he N - ~ . e ~ i n a l region .of fhe p u r i f i e d A~ pr,o~eins f ; o m B . ~t~m~othermophi~us was s.eq~eneed the N 4 e r m ~ a l a,e~idue was f o u n d t o be. rne~hi,onine za'ther than ~e~me. T h e f i r s l 1-S r e s i d u e s oi fl~e.A2 p r o t e i n o r B . s~ea.~ozl~ez~ophit~s me: Ne~-Thr- Lyz-'G]u-G~]n-Ile-1]e-Gln-Ala-N'N- L ~ - A s n - ~.e,l.... " - • .... . . Se~-?%~aL. . "Fig.2. Immanochemicai ~ , ~ i ~ :of/ft. ~t,earo~h:~moFh$~u~ ~ ' i e absen¢e o f a n . N - ~ e r m h a a l ~er~me mad the prepr,ot,eins t h a i , c o z r e ~ p . 0 n d 1.0 E. ,egli t~r0;teins L7,~L12. ,tempe " " s e n c e o f ~ : m 6 ~ h i o n i n e , a~ first ~uxpxisixag, it n o t u n l i k e we]l eonlnaned.An~ E. ,coli L12..~2) B..stearo~]J-.ezmOpBffus " th~ .da~-a r e p o t t e d f o r ithe ~ ¢ o h o l ~ e h y d r o g e n a s e f r o m A~ ~ 1 mgjma an ph:o~pha~e b~ff~rvxt saline pH 7;0 t l) 9:9% ,,sai|i:ne, ~. st,earo,'.tI~erma)pN:lus [23] w h e r e ~he ace~.y]a'ted
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F~BS L~TTERS
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M~y 1974
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!r~= ~. 4. Cerspaison o f ~h.e N-t~;m~na] arn~n~ a ~ d s~q~ences in i T - - L ] 2 f~om ~. ~ 1 i and B. s¢~ro;her.~.nop~jh'.s A2 pro~e~. "Ih~ s . ~ e n c e s a:e a~gned ~o ~ha~ ~aline-] g {~,~f. ~3] ) cor~eapongs !o ~'a~ine 15 in B. ste=zotTJe~opl_,iN~ A~. R~id~aes e n e : ~ e d by d ~ b l e lines a~e idenfi~M.
s erine is also r e p ] a e e d b y a r a e l h i o n i n e . F u r t h e r m o r e ,
F. Rollin a~d }~ Tess[ez fox expert ~e~b_nScal ~ s i s ~ c e
it .is o f in~eres~ @a~ ~he a l a n ~ e A.ch a cidi~ p~o~eins frown ihe ~xt~enae h M o p ~ o b~c~e~nrn, iz]~'!,~b~ci,e1"i,1~n
and to D~. H. Jennings m~d M~s. A. NaltSn fox help w~th the immunological experLn~.en~s. We :~ank D n
~ t i n ~ . b r a n ~ , also has ~ rnet~hionine W-~errninal (O,aa e~ al. ~ n p n b ] i s h e d o b s e r v a t i e n s ) . F o r t h e A p~ote~ns f~.om ~ co1$ a n d ~ s~e,a r ~
1-Ia_~'ey Kaplmn f ~ ~his v~/~a~ble discu~sioDs.
the~op1~ilus, ,it ~s possible
References
l:e s h o w ]~,om,o,'].ogies m the c h a i n s :f , h e m e t h i , o n m e N - t e n n m m of the/~, s 2 . e ~ t h e ~ a o p h i I u s pr,o~eh.~ ~ a]igned w i t h ~he iso].eacin,e-2 ,the ~.. c o g L 7 o~ L 1 2 p~,o~e:ns [f~g 4 ] . I f d-Bs is
done then 1i3 o u t o f 1he f i ~ t 16 ~esidu~s :(62.5%) are iden.~eal in ~la~ ,two e h ~ n s . I n t h o s e ~h~'t ,difl?~r.Tne,t1, glu-4 and gha-tg, c o u l d h a v e arisen b y poSnt rnuta~.on and the fizs,t t w o are as w.e]l, e,0nse~ad~e ehan~es.
S ~ a ~ h,omolopAes bm~e been shown fn: some 30 S p~ot,e'ms [24], Howeve:, ~he degree t.o which this h ~ molegy ~s ,~ontinU.ed in the primary strac~u~e o f th:~ A,~ and Aa pro~eh~s awa~s fu;the~ sequence ma~lyses o f these pro[eir~s :_~ B. sIe~ro.~,ermcphflus. ]n view o f ~ e homologies m primary s~ra~m:e and N - s ~ i b s ~ t u t i o n it ~s poss~Me t h a t ,theN - t e n n i n ~ l ,~e~ o n o f ~these p~ot,eins has i m p o r t a n t £uncfi.onat ~o]:es. .Ther.efo:e in ol,fl,~ t o e]uci,fla~e t h e .~ole of ,%hese p~ly~ e p f i f l e s ~ t o ~ ~ibosom~l ~une~ion i'ti~ n e c e s s a r y •t h a t t h e c h e n d e a l _~r'ope:des o f ~the amin.o t:erminns :be d e t e r m i n e & Si~eh s~u.dies h a v e n o w b e e n iNtia:e.d ,on: laboratory.
Aeknow]edgementsT h e a,i~.tho~ are i n d e b t e d ,to :C, tSoy, W; Row~o~-ne,
~1972) :E~..]. B~ochem. 2 5 . 5 - - ] 2 . ].2] T~e:;]~z-~, C-, "WitI~rsn.-I.Se~.D]d, B. a:nd ~]i~]te;, 'W. ~1'972) ~En;. 3. BSache:~.. 2 5 . 1 3 - t 9 . I 3 ] Te!hozs~, C., MSIle~, W., laursen, R. and Ni~!rnann15ebm]d, B. {1992) iFEBS ke~e~s 2g, 3 2 5 - 3 2 g . ]4] T e : h e ~ , C., M~51]~z, W., L~ursen, R. and ?,i~rnann]_SebNd, B. f1973) E~z. J. Biochrenl. 34, 13B-~52~5] Bre~, N., ~amasa.ki, E., Redfield, B. and WeSsshach. ]]. ( t 972) .A~ch. Biuchem. Bi~phys. ] 48, ] 48-- ] 55. I5] B~m, N. an~t W ~ s b a c b , H. ~]972~ Bio~hern. Biophys.
11~ C~n~mn~. 49, 673-~79. ]7] ~v'f~hsb~ch, 1-1.,~¢.d~e]d, ]3, Yamnsaki, E., D~",i~. :R. C:., P~k~, N. and Brow, N. [1972) A~c~h.~och~m. BSophys. 149, 110- 117. 1~] K~P.g, l-].-, ,, z,0x, 3. E., $pear~, C,, B~e~, ?.,'. and Weissbaeh, H. '[1973) L B]~I. Churn. 248, 5 0 ! 2 - - 5 0 t S]N} BIo't: N., Marcel R., C~ap.~, L- and W~ssb~ch, N- {1~73) A~eh. Bier-hera. Biophys. t 55: 4 7 5 - 4 7 7 . B~o=Nern. BiophN~ Res. Cornr~¢_~-~47, ~66--~873. ~ ] ~ Chin~li,O. and ]~a~/n~-g=gian~, A. '119739~-~r.3. iBieahern. 32,463 -472. I32~ Nnrnel, E. anti Nakalno'Io,~. 11972) 3. BID3. Chef-. 247, 6810-15817o i~5] l-l~rnei, E., XoLa, ~l. n~,~/~akar~.e]p, T. ~]97~2) J. ~ o l . .Chen~. 247, guZ.--B 14. P- -(a9;~.~ Biochem. 33~ophym llem Cormn'~r~ 46, 238"24~. "
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• . :.-:I.~EBS~LETTERS
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May !974
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[ i 5 ] H i # l a n d , " J..H,~ B o d t e y ~ J ~. ~#;~.; ' G o r d o n , : J ,, ~ l a s e n b a n k , ~ . . ~.[20] S r r f i t ~ O , , G i b s * n , -D~ ~F&m~ng, E , M . , G 0 o d f l i ~ h : , " a n d .$tiscfler, G . (I97~.3). :Pre-e; N ~ t f l . . A e a d . GeL U . S . 7.(i, : ' . " --: i~ R..M.~ G i l m a n , J. G . '.and B a B a m ~ n e , D., L . t 1 9 7 1 ) -~147-1$0. ~i .. " " ~:.. !.:: " • " " .- B i o d h e r r h s t r y ] 0 , 49a~-ag,2~. l~i6.1. S c h f l e r . i ' . L , M a a s s e n , ] ; . A . a n d M ~ t l e r ; W~ ( 1 9 7 3 3 ..., . ~i t21] Weber, K. andOsbom, M . ( , i 9 6 9 ) J . B i o l . C h e i n . 244~ ' . Bioclne'nxB~ophy.~ Re~ .Gommun. 53,:90--98. ..... . . --4406~.4412, |17:] C h o w , C. T..~ ~ ' i s e n t i n , L. P . , M a t h e s o n . . A . ~F; a n d _ [ 2 2 } - . . ~ ' ~/ a a . P . . K,ttri~nd. C . ;G_..Dexxgser. E - , .~a~,re~oex;J-, Yagucl~j. M . ( 1 9 7 2 3 Blochlm. Blophys. A c . a . f i g T , . • ' ~tasc~e~, ~.~, L~t~/f~'ler, G.. a~.d W i t t _ m ~ , ~]. G.(xD733 270--:28.1: - " " " • " : . - "- i . . : . . . --Na~.ure l ~ e w ~B~01. 2 4 2 , 4 7 - - 4 9 . [ 1 8 1 K a ! t s c l m ~ d : ~ , E . m ~ d w)l~m~.n.n., 14. G° ~197'~):A'na L I 2 3 ] B r i d g c n , 3 . , K o I b , ~ . a n d ~ a r f i ~ , J . I. ( 1 9 7 3 . ) : F E ] ~ S • Bi0chem:36,401-412,,. " " " i .- " ..... • I~et,t.er~BB, l - B . ..- " " [19.1 E d m a n ; P. :and BL~gg,.G. ( 1 9 6 7 ) ] { a r . J / B k , e]aem~ I , •i[24] .3?aguclfi, ~ d . , l ~ o y , C., 2VlaZhes~n~ A . T . z l a d V i s e n f i n , . g9--91~ . " L~ P. ( 1 9 7 3 ) . C a n . J - Bioclaeaz~:ti], 1 2 1 5 - - 1 2 1 7 . • .
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