Inactivation of catalytically active thermal polyanhydro-alpha-amino acids by heating in buffered solutio

Fox, S. W.

Rohlfing, D. L.

English

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T h e w l i y  prepared polyanh;ldra-a-.iPlhO 8cid0 contuiaing 
hietfaine ~mtalyS0 tho hrdmlpaim Of p-AitrOghOnll acetate. The 
ipore active preparstionr (hiatidino equivalent ba~iia) are aimoat 
toxallp izmotivatd by heating l a  buffend eolution. 
tlie of ofuiio irI40 boadr, in vhioh .#partic .aid reriduam are 
lnitlally bo-4, aeoo8p.nlor tho iaaotivation by heat. *sa of 
activity is also ob6arved whoa such io ida  bondm 8 r C  hydroiyted 
by treatment with 8 ikc .U a t  roo8 temperature. ReLate4 erperi- 
mente, both with proteiaoldr (thermal polymer. containing ail 
aaino acid8 eonon t o  protola) and C O n r p O O i t i O M l l y  s imple co- 
polymers of arputic mold d hirtidlao, indiaato that the 
sirultaceous premen138 in the ram McrOBo~ecruiU prepsx?atioa o f  
hlrtldlne reai4ues and imide  link450r of asp.rtio acid reaiffucrr 
The hgbroip 
I s  essentiai for the higher lorelo of act iv i ty  obr@rrchf. 
Po~paahydro-cl-arino acids contslniryC the 8id;bto.n amino aci38 
comma t o  protein (protolaold8) ara proparod by h~atiag the proper 
proportions of initially dry amino acid8 (i and biblio@?sphy). 
Arow the b.tl;l proportlor that proteiaoldr oxhibit in c o m n  w i t h  
protein6 I8 that of oatalytio aotsvity (2-10)D A prorloua report 
( 2 )  ha8 r O V 8 a c d  th8f p m t O i m i b @  aafrlYS0 tho ) p r b r O l j r O i 8  Of 2- 
nftrophrayl aoot.te ( I I I P A ) ,  that himtidine roridu88 play a koy 
role ln ?he o a f a l p l a ,  and that mme proteinold8 axw m a n y  tire6 
as 8ctlre (up t o  15 t i roo)  am tho oqulvalent Uount of f r e e  
hi8tIdiP41, 886Uitr Of tho pE888Xlt inV.8tig.tiOn ahow that t h e  
more eotire protainoib8 o m  be aimoat totally imctfv8t.d by hoat- 
- 2 -  
- 3 -  
I -  
# 
- 4 -  
RESULTS MiD DISCUSSIOE 
The oatairtio action on &PA W the iMcrtintioa by heat of 
008 proteinoid -0 depiotod la P i a .  I. A@ ovaluatod fro8 tho 
- 5 -  
TABU I: 
3.m 
0.60 
0.72 
0088 
1.10 
Om70 
Om36 
0.39 
Om54 
0.05 
Om26 
0.01 
ib10 
0.50 
Om96 
l a 0 8  
Om48 
2-32 
2.77 
Om00 
3 . d  
w38= 
0.14 
0.05 
0. 42 
0.70 
0.98 
Om 22 
0.12 
0.54 
OD35 
0.25 
-0.01 
- 
Om50 
Om38 
Om72 
Om70 
0.16 
l a 7 3  
2.25 
7 
5.45: 
3 -85 
95 
92 
40 
20 
il 
69 
69 
37 
58 
4 
- 
- 
55 
24 
25 
f 5  
62 
25 
19 - 
1 
1 
- 6 -  . 
- 7 -  
Is attributed t o  amido (poptido? 23). Similar experfnoate h8ve 
- 8 -  
I Table XI &0 troor6.d the result. of an axperirent i n  vhich tho 
d i e l y s i s  a d  IpQphiliWtiOIL 8 t e p l  ~8rpB omitted, and l a  *%oh r C t l -  
r l t y  on H P A  and speaif ic  absorptivity of the prront polymer vera 
s v a b a t d  in r ~ t s n e o u 8  corprari6on vifh those of the heated, base- 
t rmtad ,  Qpd bare-trestad-aad-hertidl en8p1obe The 8psC?ifiU 
absorptivitier indlcate that either heat- in buffer or treating 
with alkali a t  room temperature ramal%. In the .care degreo of 
OpeZLhg Of Wid.; O l t h O r  fc.at8Ortt a8 8qUrlly deWh8Atal  t 0  the 
c3ataiytio sct l r l ty .  Heatlag the W e - t r e a t e d  (fee. Imide-free) 
sample d i 3  not migaffirrantly affect either property. 
remalte, obtained with oopolymer of a8partic aoid and histiaine, 
i l l U O t M ? X  the hportracs of imido, ond bec8-e the-1 poly- 
aopartio m i d  (oont.iniag imido but m him din^) i~ iaaotire 
(Tabre I), one may o o ~ l u d e  that the 8isulta~ieou8 prorenos in the 
mmc maamwlaauier preparation of bath hlrtidiae reridues and 
iaide ligkagem of 8 8 p U f I o  acid remiduer 1. asaentlel for an en- 
hanotd ierei of sotiri:y. 
These 
In experiments in rhioh rarplos of polmar WPO MI0v.d f r o m  
tho Tltrlgraph after. rUrious ext8nt8 of by-1y.i. of %aide, %t 
ua8 found that the degreo of crtrlrfict ratirity -8 fn put 
directiy proportional to t h e  mot ta t  of imido pmaont In the poly- 
mer (1-2.8-b), w i t h  mono 8otfrit7 rc#cl;iniag rftor t o f a  0p.rring 
o f  h i d @ .  A sirllar r e r u l t  wu8 obtalned when irldm n m  fmatian- 
a l l y  opened by heatlag for  15 .la. a t  differoat tapantur8. 
(Sb-lOO*C) e 
Poasiblo way8 by uhleh the h i d @  lhk-8 n d i a t o  it6 
effmct n y  Include the followi-t Tho grasr ooaforUtion o f  the 
- 9 -  
I TABLE 11 
byaluea in parentheres a r e  m l a t l t e  to the (untreated) 
oantrol. 

- 11 - 
4 
b i d 8  bonds. 
‘;‘here co#sidsrattoas ra ia tr  t o  prroteinofrft~ a. oodeis of primi-  
t i V e  abiotia protein (1.3.31). 
by protaiaoidrs suggeot how primitive protein-like macronolecules 
vith inactiratabh cratautic povmr oould hare been formed abio- 
genica iiy from amino acidr. 
Them pud other re#rlt. on Cat&3iy818 
? - 12 - 
i i 
3.. 
2. 
I 
3 0  
4 .  
5. 
6 .  
7- 
Om 
9 .  
10. 
i l e  
12 0 
13. 
14. 
13 . 
16 0 
i7 0 
18 . 
19 0 
20 0 
 OX, 2 ,  k., h'atwa, a, 328 (1965). 
- 1) - 
f 
I- 
w 
I 
Z 
3 
a 
v) 
3 
0 w z 
Q: I- 
O 
0 
... 
I -  , -  -.. 
- i4 - 
F i ~ a  1. Effeet of boating irr buff8r O'II the mtlrity of 
proteinoid A-2.8. 
a8 deaarlbed tn the ItBTHODS ~oetioa, except th.t 
tht pX n 8  6.2. Hoaffllg tho buffet aioa8 hQd no 
di8comiblo offoot On the rate of 8po&tua.auu 
l@rolymim 
The oonditionr of aamy wre 

- 15 - 
Figure 2. Ultraviolet abrrorption opectr8 of polymer 8-2.8 
before .Dd aft8r haatlag iP buffer or trutimg 
with rUall. C o n c o ~ t r a t l o ~  WZQ 8.8 mg./l. 
in 0,067 lul phoaphato buffer, pfI 6.6. Buffor 
alkali-fraatsd ample n o  not slgniflaurfly 
-8 U r d  M the bl-. The r p O t -  Of tb 
8ffaot.h h0.t-e 


The Inactivation of Catalytically Active Thermal Polyanhydro-alpha-amino Acids. /inactivation of Thermal Polyamino Acids/

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