The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3

Troeberg, Linda, Fushimi, Kazunari, Scilabra, Simone D, Nakamura, Hiroyuki, Dive, Vincent, Thøgersen, Ida B, Enghild, Jan J and Nagase, Hideaki (2009) The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3. Matrix Biology, 28 (8). pp. 463-469. ISSN 0945-053X

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Abstract

We investigated whether the affinity of tissue inhibitor of metalloproteinases (TIMP)-3 for adamalysins with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 is affected by the non-catalytic ancillary domains of the enzymes. For this purpose, we first established a novel method of purifying recombinant FLAG-tagged TIMP-3 and its inhibitory N-terminal domain (N-TIMP-3) by treating transfected HEK293 cells with sodium chlorate to prevent heparan sulfate proteoglycan-mediated TIMP-3 internalization. TIMP-3 and N-TIMP-3 affinity for selected matrix metalloproteinases and forms of ADAMTS-4 and -5 lacking sequential C-terminal domains was determined. TIMP-3 and N-TIMP-3 displayed similar affinity for various matrix metalloproteinases as has been previously reported for E. coli-expressed N-TIMP-3. ADAMTS-4 and -5 were inhibited more strongly by N-TIMP-3 than by full-length TIMP-3. The C-terminal domains of the enzymes enhanced interaction with N-TIMP-3 and to a lesser extent with the full-length inhibitor. For example, N-TIMP-3 had 7.5-fold better K(i) value for full-length ADAMTS-5 than for the catalytic and disintegrin domain alone. We propose that the C-terminal domains of the enzymes affect the structure around the active site, favouring interaction with TIMP-3.

Item Type: Article
Uncontrolled Keywords: antagonists & inhibitors,adamts4 protein,adamts5 protein,drug effects,cell line,glycosylation,humans,kinetics,genetics,genetics,genetics,matrix metalloproteinase inhibitors,antagonists & inhibitors,physiology,physiology,biosynthesis,biosynthesis,transfection
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User: LivePure Connector
Date Deposited: 08 Jan 2019 12:30
Last Modified: 22 Apr 2020 07:22
URI: https://ueaeprints.uea.ac.uk/id/eprint/69490
DOI: 10.1016/j.matbio.2009.07.005

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