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FEBS Lett. 1995 Feb 20;360(1):52-6.

Synthesis and refolding of human TIMP-2 from E. coli, with specific activity for MMP-2.

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1
Department of Biochemistry, School of Veterinary Medicine, University of Padova, Italy.

Abstract

Tissue inhibitors of metalloproteinase (TIMPs) are inhibitory counterparts of collagenases, containing 12 cysteine residues paired to six internal disulphide bridges. TIMP-2, an inhibitory protein of 72 kDa gelatinase/type IV collagenase (MMP-2), was expressed in Escherichia coli as a fusion protein with a 34 amino acid NH2-linked tail containing six consecutive histidine residues. The protein was purified in a single-step using an ion metal affinity column (IMAC) in denaturing conditions. The immobilized fusion TIMP-2 protein was refolded at a high concentration in the column, producing about 5 mg of protein per litre of bacterial cells. It shows specific binding and inhibitory activity against MMP-2, but has no effect against 92 and 45 kDa gelatinases.

PMID:
7875301
DOI:
10.1016/0014-5793(95)00073-i
[Indexed for MEDLINE]
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