2 SLOW STAGES IN REFOLDING OF BOVINE CARBONIC ANHYDRASE-B ARE DUE TO PROLINE ISOMERIZATION

被引:46
作者
SEMISOTNOV, GV [1 ]
UVERSKY, VN [1 ]
SOKOLOVSKY, IV [1 ]
GUTIN, AM [1 ]
RAZGULYAEV, OI [1 ]
RODIONOVA, NA [1 ]
机构
[1] ACAD SCI USSR,INST PROT RES,PUSHCHINO 142292,USSR
关键词
D O I
10.1016/S0022-2836(05)80215-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Kinetics of refolding of bovine carbonic anhydrase B have been studied by the "double-jump" technique (i.e. the dependence of protein refolding on delay time in the unfolded state after fast unfolding). It is shown that two stages (the slow with a relaxation time of t1/2 ≈ 120 s and the superslow with t1/2 ≈ 600 s) observed during refolding of bovine carbonic anhydrase B are due to trans-cis isomerization of proline residues. The dependences of rate constants of these processes on temperature and on the final denaturant concentration were measured. Activation energies of both processes are the same, Ea = 18(±2) kcal/mol. The rate constants of protein refolding do not depend on the final concentration of urea under native conditions. In addition, the rate of isomerization of essential proline residues in the "molten globule" intermediate state of bovine carbonic anhydrase was measured and found to be equal to that for unstructural polypeptides. The effect of several proline residues on carbonic anhydrase refolding is discussed. © 1990 Academic Press Limited.
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页码:561 / 568
页数:8
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