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Cloning of rat 92‐kDa type IV collagenase and expression of an active recombinant catalytic domain
Author(s) -
Xia Yiyang,
Garcia Gabriela,
Chen Shizhong,
Wilson Curtis B.,
Feng Lili
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00185-8
Subject(s) - recombinant dna , cloning (programming) , collagenase , microbiology and biotechnology , expression cloning , chemistry , molecular cloning , biology , biochemistry , gene expression , complementary dna , gene , enzyme , computer science , programming language
A full‐length cDNA for rat 92‐kDa type IV collagenase was isolated and sequenced. RNase protection assay revealed tissue specific differential expression of the 92‐kDa type IV collagenase in the rat during development. Natural and modified forms of the 92‐kDa type IV collagenase were expressed. One active protein, 92‐CD, contained only the putative catalytic domain. Large quantities of the 92‐CD were expressed in Escherichia coli , extracted from inclusion bodies, purified, and refolded to an active form. This recombinant protein was able to cleave denatured and native collagen and was inactivated by known MMP inhibitors.