Open AccessMolecular cloning and functional characterization of a snake venom metalloprotease
Author(s) -
Jeon OkHee,
Kim DooSik
Publication year - 1999
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1999.00525.x
Subject(s) - venom , metalloproteinase , snake venom , cloning (programming) , computational biology , molecular cloning , disintegrin , biology , matrix metalloproteinase , genetics , gene , biochemistry , computer science , peptide sequence , programming language
A cDNA clone, MT‐d, encoding metalloprotease precursor was isolated from snake ( Agkistrodon halys brevicaudus ) venom gland cDNA library. MT‐d‐I protein containing both metalloprotease and disintegrin domains, and MT‐d‐II protein containing the metalloprotease domain only were expressed in Escherichia coli and refolded successfully into their functional forms. Each of the refolded enzyme species exhibited distinct substrate specificity. Proteolytic activity of the MT‐d‐1 was able to hydrolyse type I gelatin, type‐III and V collagens in contrast with the catalytic function of MT‐d‐II. MT‐d‐I protein having metalloprotease activity was also able to inhibit platelet aggregation. Functionally active MT‐d‐I protein underwent autoproteolytic processing in vitro to produce metalloprotease and disintegrin; this processing was accompanied by significant changes in the substrate specificity of the enzyme activity. Experimental evidence strongly suggests that the disintegrin domain in the metalloprotease precursor modulates the catalytic function of the enzyme in hydrolysing extracellular matrix proteins.