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A disintegrin and metalloproteinase with thrombospondin motifs - 5 (ADAMTS - 5) forms catalytically active oligomers

Citation

Kosiah, HJ and Last, K and Rogerson, FM and Golub, SB and Gauci, SJ and Russo, VC and Stanton, H and Wilson, RR and Lamamde, SR and Holden, P and Fosang, AJ, A disintegrin and metalloproteinase with thrombospondin motifs - 5 (ADAMTS - 5) forms catalytically active oligomers, Journal of Biological Chemistry, 291, (7) pp. 3197-3208. ISSN 1083-351X (2016) [Refereed Article]


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Copyright Statement

Copyright 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

DOI: doi:10.1074/jbc.M115.704817

Abstract

The metalloproteinase ADAMTS-5 (A disintegrin and metalloproteinase with thrombospondin motifs) degrades aggrecan, a proteoglycan essential for cartilage structure and function. ADAMTS-5 is the major aggrecanase in mouse cartilage, and is also likely to be the major aggrecanase in humans. ADAMTS-5 is a multidomain enzyme, but the function of the C-terminal ancillary domains is poorly understood. We show that mutant ADAMTS-5 lacking the catalytic domain, but with a full suite of ancillary domains inhibits wild type ADAMTS activity, in vitro and in vivo, in a dominant-negative manner. The data suggest that mutant ADAMTS-5 binds to wild type ADAMTS-5; thus we tested the hypothesis that ADAMTS-5 associates to form oligomers. Co-elution, competition, and in situ PLA experiments using full-length and truncated recombinant ADAMTS-5 confirmed that ADAMTS-5 molecules interact, and showed that the catalytic and disintegrin-like domains support these intermolecular interactions. Cross-linking experiments revealed that recombinant ADAMTS-5 formed large, reduction-sensitive oligomers with a nominal molecular mass of ∼400 kDa. The oligomers were unimolecular and proteolytically active. ADAMTS-5 truncates comprising the disintegrin and/or catalytic domains were able to competitively block full-length ADAMTS-5-mediated aggrecan cleavage, measured by production of the G1-EGE373 neoepitope. These results show that ADAMTS-5 oligomerization is required for full aggrecanase activity, and they provide evidence that blocking oligomerization inhibits ADAMTS-5 activity. The data identify the surface provided by the catalytic and disintegrin-like domains of ADAMTS-5 as a legitimate target for the design of aggrecanase inhibitors.

Item Details

Item Type:Refereed Article
Keywords:ADAMTS, ADAMTS5, aggrecan, cartilage, cartilage biology, metalloprotease, oligomer, oligomerization
Research Division:Biological Sciences
Research Group:Biochemistry and Cell Biology
Research Field:Enzymes
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Skeletal System and Disorders (incl. Arthritis)
Author:Wilson, RR (Dr Richard Wilson)
ID Code:106455
Year Published:2016 (online first 2015)
Web of Science® Times Cited:3
Deposited By:Central Science Laboratory
Deposited On:2016-02-10
Last Modified:2017-11-07
Downloads:12 View Download Statistics

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