Expression of rat chymotrypsinogen in yeast: a study on the structural and functional significance of the chymotrypsinogen propeptide

FEBS Letters

Rutter, William J.²  Gráf, László¹  Venekei, István² 

[1] Department of Biochemistry, Eotvos University, Puskin ut. 3, Budapest, H-1088, Hungary;Hormone Research Institute, University of California San Francisco, San Francisco, CA 94143-0534, USA

关键词: Zymogen activation;    Chymotrypsin;    Disulfide bond;    Protein folding;    TPCK;    tosyl-l-alanyl chloromethyl ketone;    SBTI;    soybean trypsin inhibitor;    pNA;    p-nitroanilide;    AMC;    7-amino-4-methylcoumarin;    MUGB;    4-methylumbelliferyl p-guanidinobenzoate;    MUTMAC;    4-methylumbelliferyl p-(N N;    N-trimethyl ammonium) cinnamate chloride;   

DOI  :  10.1016/0014-5793(95)01483-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The role of the propeptide sequence and a disulfide bridge between sites 1 and 122 in chymotrypsin has been examined by comparing enzyme activities of wild-type and mutant enzymes. The kinetic constants of mutants devoid of the Cys1-Cys122 disulfide-linked propeptide show that this linkage is not important either for activity or substrate specificity. However this linkage appears to be the major factor in keeping the zymogen stable against non-specific activation. A comparison of zymogen stabilities showed that the trypsinogen propeptide is ten times more effective than the chymotrypsinogen propeptide in preventing non-specific zymogen activation during heterologous expression and secretion from yeast. This feature can also be transferred in trans to chymotrypsinogen; i.e. the chymotrypsin trypsin propeptide chimera forms a stable zymogen.

【 授权许可】

Unknown