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Cloning, expression, and characterization of a cDNA encoding snake venom metalloprotease
Author(s) -
Jeon OkHee,
Kim DooSik
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201443
Subject(s) - disintegrin , metalloproteinase , complementary dna , microbiology and biotechnology , snake venom , biochemistry , peptide sequence , recombinant dna , biology , signal peptide , protease , molecular cloning , cdna library , venom , amino acid , enzyme , gene
A cDNA clone, MT‐c, encoding metalloprotease was isolated from snake (Agkistrodon halys brevicadus) venom gland cDNA library. Deduced amino acid sequence indicated that MT‐c is composed of a signal sequence, amino‐terminal propeptide, a central metalloprotease domain, and a Lys‐Gly‐Asp (KGD) disintegrin domain. The partial cDNA encoding metalloprotease and disintegrin domain was subcloned and expressed in E. coll. The expressed MT‐c protein was purified and successfully refolded into functional form retaining the enzyme activity. Analyses of the purified recombinant protease activity revealed that the enzyme hydrolyzes extraceUular matrix proteins including type I gelatin, type 1V and type V collagen, while type I, II, III collagens and fibronectin were insensitive to the proteolytic digestion. The recombinant enzyme was also able to degrade fibrinogen by specifically cleaving Aα chain of the protein.