Site-Directed Mutations That Alter the Inhibitory Activity of the Tissue Inhibitor of Metalloproteinases-1 - Importance of the N-Terminal Region between Cysteine-3 and Cysteine-13

Oshea, Mark, Willenbrock, Frances, Williamson, Richard A., Cockett, Mark I., Freedman, Robert B., Reynolds, John J., Docherty, Andrew J. P., Murphy, Gillian (1992) Site-Directed Mutations That Alter the Inhibitory Activity of the Tissue Inhibitor of Metalloproteinases-1 - Importance of the N-Terminal Region between Cysteine-3 and Cysteine-13. Biochemistry, 31 (42). pp. 10146-10152. ISSN 0006-2960. (doi:10.1021/bi00157a002) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:22387)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.
Official URL: http://dx.doi.org/10.1021/bi00157a002

Abstract

The tissue inhibitor of metalloproteinases-1 (TIMP-1) was subjected to single-site mutations within the N-terminal three loops using an oligonucleotide-directed polymerase chain reaction method. All the histidines, and a number of other residues conserved between TIMP-1 and TIMP-2, were individually modified and the mutant TIMPs expressed in mammalian cells. Purified mutant TIMPs were shown to be correctly folded by measuring the effect of guanidine hydrochloride on intrinsic fluorescence. Kinetic analyses of mutants using a quenched fluorescent peptide substrate and the metalloproteinase PUMP indicated that mutation of His7 and Gln9 caused an increase in the apparent dissociation constant, largely due to an increase in the rate of dissociation of complexes. The data indicate that the anchored sequence between Cys 3 and Cys 13 is a key region for interaction of TIMP-1 with metalloproteinases.

Item Type:	Article
DOI/Identification number:	10.1021/bi00157a002
Subjects:	Q Science > QD Chemistry
Divisions:	Divisions > Division of Natural Sciences > Biosciences
Depositing User:	M. Nasiriavanaki
Date Deposited:	23 Aug 2009 15:10 UTC
Last Modified:	05 Nov 2024 10:01 UTC
Resource URI:	https://kar.kent.ac.uk/id/eprint/22387 (The current URI for this page, for reference purposes)

University of Kent Author Information

Williamson, Richard A..

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