Williamson, Richard A., Smith, Bryan J., Angal, Sarojani, Freedman, Robert B. (1993) Chemical modification of tissue inhibitor of metalloproteinases-1 and its inactivation by diethyl pyrocarbonate. Biochimica Et Biophysica Acta, 1203 (1). pp. 147-154. ISSN 0006-3002. (doi:10.1016/0167-4838(93)90049-w) (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:20741)
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: https://doi.org/10.1016/0167-4838(93)90049-w |
Abstract
Tissue inhibitor of metalloproteinases-1 (TIMP-1) was treated with a range of chemical modification reagents in order to identify amino acid residues essential for inhibitory activity. Diethyl pyrocarbonate (DEPC) was found to be a potent inactivator at low reagent/TIMP molar concentrations. The extent of modification at 50% inactivation was determined as 1.5 sites/molecule. The DEPC-modified inhibitor did not form stable complexes with stromelysin, but was shown to retain native structure as judged by conformational stability to denaturation by guanidine hydrochloride. Peptide mapping experiments were used to find the sites of DEPC incorporation within the primary structure of TIMP and three residues were identified (His-95, His-144 and His-164). Mutant TIMPs in which histidine residues have been substituted or deleted retain inhibitory activity and were found to be equally as sensitive to DEPC inactivation as the wild-type. No new sites of DEPC modification in the mutant proteins were detected. The possible contribution made by His residues 95, 144 and 164 to the inhibitory activity of TIMP is discussed.
Item Type: | Article |
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DOI/Identification number: | 10.1016/0167-4838(93)90049-w |
Uncontrolled keywords: | Timp; tissue inhibitor; metalloproteinase; chemical modification; diethyl pyrocarbonate; peptide mapping |
Subjects: |
Q Science > QP Physiology (Living systems) > QP517 Biochemistry Q Science > QP Physiology (Living systems) > QP506 Molecular biology |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | O.O. Odanye |
Date Deposited: | 13 Jul 2009 19:43 UTC |
Last Modified: | 09 Mar 2023 11:31 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/20741 (The current URI for this page, for reference purposes) |
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