Cloning, expression, and characterization of a cDNA encoding snake venom metalloprotease.
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. coli. 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 extracellular matrix proteins including type I gelatin, type IV 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 A alpha chain of the protein.
Jeon
O H
OH
Department of Biochemistry, College of Science, Yonsei University, Seoul, Korea.
Kim
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DS
D017836:Agkistrodon / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000201:enzymology*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D007723:Korea; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D017510:Protein Folding; D013379:Substrate Specificity
D003435:Crotalid Venoms; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D005340:Fibrinogen; D008666:Metalloendopeptidases