Activation of snake venom metalloproteinases by a cysteine switch‐like mechanism

FEBS Letters

Huber, Robert²  Grams, Frank²  Bode, Wolfram²  Moroder, Luis²  Kress, Lawrence F.¹ 

[1] Department of Molecular and Cellular Biology, Roswell Park Cancer Institute. Buffalo, NY 14263, USA;Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, 82152 Martinsried, Germany

关键词: Cysteine switch;    Matrix metalloproteinase;    Snake venom metalloproteinase;    Adamalysin;    Proteinase activation;    Stability of cysteine peptide;    standard abbreviations for amino acids and peptide derivatives are used as recommended by the IUPAC-IUB commission on biochemical nomenclature. The amino acids are of l-configuration unless stated otherwise. HPLC;    high-pressure liquid chromatography;    FAB-MS;    fast atom bombardment mass spectroscopy;    Abz;    2-aminobenzoyl;   

DOI  :  10.1016/0014-5793(93)80443-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNAs of several snake venom zinc endopeptidases code for a putative propeptide, which includes the conserved cysteine-containing sequence PKMCGVT. It has been suggested that binding of the cysteine thiol function to the active-site zinc, resulting in inactivation of the catalytic domain, occurs in a mode similar to the ‘cysteine switch’ mechanism proposed for matrix metalloproteinases. In order to confirm this hypothesis, inhibition kinetics have been performed on the metalloproteinase adamalysin II of the venom of the snake Crotalus adamanteus using several cysteine peptides. Among these the synthetic hexapeptide PKMCGV-NH₂, corresponding to the conserved sequence portion of the known propeptides, was found to be by far the strongest inhibitor of this proteinase with a K _i of 3.4 μM. The inhibitory potencies of an equivalent peptide with the l-Cys replaced by a d-Cys or by an l-Ser as well as of reduced glutathione, cysteine and two unrelated cysteine peptides were by one to two orders of magnitudes lower. These findings strongly support a cysteine switch-like mechanism even for activation of the snake venom metalloproteinases.

【 授权许可】

Unknown