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

Troeberg, Linda ORCID: https://orcid.org/0000-0003-0939-4651, 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
UEA Research Groups:	Faculty of Medicine and Health Sciences > Research Groups > Musculoskeletal Medicine Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health
Depositing User:	LivePure Connector
Date Deposited:	08 Jan 2019 12:30
Last Modified:	19 Oct 2023 02:20
URI:	https://ueaeprints.uea.ac.uk/id/eprint/69490
DOI:	10.1016/j.matbio.2009.07.005

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