SOME KINETIC-PROPERTIES OF A CYSTEINE PROTEINASE (CRUZIPAIN) FROM TRYPANOSOMA-CRUZI

被引:142
作者
CAZZULO, JJ
FRANKE, MCC
MARTINEZ, J
DECAZZULO, BMF
机构
[1] Instituto de Investigaciones Bioquimicas Fundación Campomar, Universidad de Buenos Aires-CONICET, Buenos Aires
关键词
(Trypanosoma cruzi); Cruzipain; Cysteine proteinase; Substrate specificity; Urea activation;
D O I
10.1016/0167-4838(90)90166-D
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
A cysteine proteinase, purified to homogeneity from epimastigotes of Trypanosoma cruzi, was strongly inhibited by l-trans-epoxysuccinylleucylamido(4-guanidino)butane (E-64). The second-order rate constant was 20 800 M-1·s-1, and the reagent could be used for active site titration. The enzyme hydrolysed chromogenic peptides at the carboxyl Arg or Lys; it required at least one more amino acid, preferably Arg, Phe, Val or Leu, between the terminal Arg or Lys and the amino-blocking group. Enzyme activity on azocasein at pH 5.0 was increased by urea, maximal activity being attained at 2 M, and was still as active at 5 M urea as in its absence. Guanidine hydrochloride and KSCN also activated at low concentrations, but caused a strong inhibition above 2 M and 1 M, respectively. When azocasein was tested as a substrate at pH 7.0, there was no activation, and when synthetic substrates were used all chaotropic agents tested were inhibitory. The results suggest that the enzyme, for which we propose the trivial name 'cruzipain', differs in some aspects from all other cysteine proteinases described so far, although it shares several of the properties of mammalian cathepsin L. © 1990.
引用
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页码:186 / 191
页数:6
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