106455 - A disintegrin and metalloproteinase.pdf (2.46 MB)
A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) forms catalytically active oligomers
journal contribution
posted on 2023-05-18, 16:48 authored by Kosiah, HJ, Last, K, Rogerson, FM, Golub, SB, Gauci, SJ, Russo, VC, Stanton, H, Richard WilsonRichard Wilson, Lamamde, SR, Holden, P, Fosang, AJThe 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.
History
Publication title
Journal of Biological ChemistryVolume
291Issue
7Pagination
3197-3208ISSN
1083-351XPublisher
American Society of Biochemistry and Molecular BiologyPlace of publication
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Copyright 2016 by The American Society for Biochemistry and Molecular Biology, Inc.Repository Status
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