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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, R 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 |
| Objective Field: | Clinical health not elsewhere classified |
| UTAS Author: | Wilson, R (Dr Richard Wilson) |
| ID Code: | 106455 |
| Year Published: | 2016 (online first 2015) |
| Web of Science® Times Cited: | 8 |
| Deposited By: | Central Science Laboratory |
| Deposited On: | 2016-02-10 |
| Last Modified: | 2021-02-18 |
| Downloads: | 125 View Download Statistics |
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