Quantitative proteomics reveals that ADAM15 can have proteolytic-independent functions in the steady state

Yang, Chun Yao, Bonelli, Simone, Calligaris, Matteo, Carreca, Anna Paola, Müller, Stephan A., Lichtenthaler, Stefan F., Troeberg, Linda ORCID: https://orcid.org/0000-0003-0939-4651 and Scilabra, Simone D. (2022) Quantitative proteomics reveals that ADAM15 can have proteolytic-independent functions in the steady state. Membranes, 12 (6). ISSN 2077-0375

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A disintegrin and metalloproteinase 15 (ADAM15) is a member of the ADAM family of sheddases. Its genetic ablation in mice suggests that ADAM15 plays an important role in a wide variety of biological functions, including cartilage homeostasis. Nevertheless, while the substrate repertoire of other members of the ADAM family, including ADAM10 and ADAM17, is largely established, little is known about the substrates of ADAM15 and how it exerts its biological functions. Herein, we used unbiased proteomics to identify ADAM15 substrates and proteins regulated by the proteinase in chondrocyte-like HTB94 cells. ADAM15 silencing did not induce major changes in the secretome composition of HTB94 cells, as revealed by two different proteomic approaches. Conversely, overexpression of ADAM15 remodeled the secretome, with levels of several secreted proteins being altered compared to GFP-overexpressing controls. However, the analysis did not identify potential substrates of the sheddase, i.e., transmembrane proteins released by ADAM15 in the extracellular milieu. Intriguingly, secretome analysis and immunoblotting demonstrated that ADAM15 overexpression increased secreted levels of tissue inhibitor of metalloproteinases 3 (TIMP-3), a major regulator of extracellular matrix turnover. An inactive form of ADAM15 led to a similar increase in the inhibitor, indicating that ADAM15 regulates TIMP-3 secretion by an unknown mechanism independent of its catalytic activity. In conclusion, high-resolution quantitative proteomics of HTB94 cells manipulated to have increased or decreased ADAM15 expression did not identify canonical substrates of the proteinase in the steady state, but it revealed that ADAM15 can modulate the secretome in a catalytically-independent manner.

Item Type: Article
Additional Information: Funding Information: Funding: This study was supported by the Kennedy Trust for Rheumatology Research. S.F.L. was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy—ID 390857198). S.D.S. was supported by Fondazione con il Sud under the Programme “Brains to South 2018”—Project “i-Rhom2: a new therapeutic target in osteoarthritis” (Grant Agreement No. 2018—PDR—00799).
Uncontrolled Keywords: adam15,adams,metalloproteinases,proteomics,secretome,chemical engineering (miscellaneous),process chemistry and technology,filtration and separation ,/dk/atira/pure/subjectarea/asjc/1500/1501
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
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Depositing User: LivePure Connector
Date Deposited: 10 Aug 2022 08:38
Last Modified: 19 Oct 2023 03:24
URI: https://ueaeprints.uea.ac.uk/id/eprint/87160
DOI: 10.3390/membranes12060578


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