EVIDENCE SUGGESTING THAT SOME PROTEOLYTIC-ENZYMES MAY CLEAVE ONLY THE TRANS FORM OF THE PEPTIDE-BOND

被引:74
作者
LIN, LN [1 ]
BRANDTS, JF [1 ]
机构
[1] UNIV MASSACHUSETTS,DEPT CHEM,AMHERST,MA 01003
关键词
D O I
10.1021/bi00568a007
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
The rates of hydrolysis of glycyl-l-proline and l-phenylalanyl-l-proline, catalyzed by prolidase, have been measured at several temperatures under conditions where a high ratio of prolidase activity to substrate concentration existed. Two well-separated kinetic phases, which can be adequately treated as two first-order reactions, were observed for the hydrolysis. The relative amplitudes of the two phases are nearly independent of temperature, but strongly dependent on the initial state of protonation of the dipeptides. It was found that the amplitude of the slow phase is strictly proportional to the known amount of cis isomer, while the amplitude of the fast phase correlates with the amount of the trans isomer. Furthermore, the relaxation time and activation energy of the slow phase of hydrolysis are in good agreement with the same parameters determined for cis-trans isomerization of the dipeptides, as measured by a pH-jump method for samples not being hydrolyzed. These results lead us to the conclusion that the slow phase seen for hydrolysis is rate limited by cis-trans isomerization of the X-Pro peptide bond. Thus, this proline-specific protease appears to have an absolute requirement for the trans form of the peptide bond and appears not to cleave the cis form or to cleave it extremely slowly. © 1979, American Chemical Society. All rights reserved.
引用
收藏
页码:43 / 47
页数:5
相关论文
共 19 条
[1]   CONSIDERATION OF POSSIBILITY THAT SLOW STEP IN PROTEIN DENATURATION REACTIONS IS DUE TO CIS-TRANS ISOMERISM OF PROLINE RESIDUES [J].
BRANDTS, JF ;
HALVORSON, HR ;
BRENNAN, M .
BIOCHEMISTRY, 1975, 14 (22) :4953-4963
[2]   UNFOLDING AND REFOLDING OCCUR MUCH FASTER FOR A PROLINE-FREE PROTEIN THAN FOR MOST PROLINE-CONTAINING PROTEINS [J].
BRANDTS, JF ;
BRENNAN, M ;
LIN, LN .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (10) :4178-4181
[3]   CONFORMATION STATES OF CONCANAVALIN A - KINETICS OF TRANSITIONS INDUCED BY INTERACTION WITH MN2+ AND CA2+ IONS [J].
BROWN, RD ;
BREWER, CF ;
KOENIG, SH .
BIOCHEMISTRY, 1977, 16 (17) :3883-3896
[4]   CIS-TRANS EQUILIBRIUM AND KINETIC STUDIES OF ACETYL-L-PROLINE AND GLYCYL-L-PROLINE [J].
CHENG, HN ;
BOVEY, FA .
BIOPOLYMERS, 1977, 16 (07) :1465-1472
[5]  
DAVIS NC, 1957, J BIOL CHEM, V224, P261
[6]   NUCLEAR MAGNETIC-RESONANCE STUDIES OF ACID-BASE CHEMISTRY OF AMINO-ACIDS AND PEPTIDES .2. DEPENDENCE OF ACIDITY OF C-TERMINAL CARBOXYL GROUP ON CONFORMATION OF C-TERMINAL PEPTIDE-BOND [J].
EVANS, CA ;
RABENSTEIN, DL .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1974, 96 (23) :7312-7317
[7]   INTERCONVERSION OF CONFORMATIONAL ISOMERS OF LIGHT-CHAINS IN MCG IMMUNOGLOBULINS [J].
FIRCA, JR ;
ELY, KR ;
KREMSER, P ;
WESTHOLM, FA ;
DORRINGTON, KJ ;
EDMUNDSON, AB .
BIOCHEMISTRY, 1978, 17 (01) :148-158
[8]   NMR-STUDIES OF MOLECULAR-CONFORMATIONS IN LINEAR OLIGOPEPTIDES H-(L-ALA)N-L-PRO-OH [J].
GRATHWOHL, C ;
WUTHRICH, K .
BIOPOLYMERS, 1976, 15 (10) :2043-2057
[9]   X-PRO PEPTIDE-BOND AS AN NMR PROBE FOR CONFORMATIONAL STUDIES OF FLEXIBLE LINEAR PEPTIDES [J].
GRATHWOHL, C ;
WUTHRICH, K .
BIOPOLYMERS, 1976, 15 (10) :2025-2041
[10]   INTESTINAL DIPEPTIDASES .I. SPECTROPHOTOMETRIC DETERMINATION AND CHARACTERIZATION OF DIPEPTIDASE ACTIVITY IN PIG INTESTINAL MUCOSA [J].
JOSEFSSON, L ;
LINDBERG, T .
BIOCHIMICA ET BIOPHYSICA ACTA, 1965, 105 (01) :149-+