Development of a fluorogenic ADAMTS-7 substrate
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Salvatore Santamaria Frederic Buemi Elisa Nuti Doretta Cuffaro Elena De Vita Tiziano Tuccinardi Armando Rossello Steven Howell Shahid Mehmood Bram Snijders Rens de GrootAbstract
The extracellular protease ADAMTS-7 has been identified as a potential therapeutic target in atherosclerosis and associated diseases such as coronary artery disease (CAD). However, ADAMTS-7 inhibitors have not been reported so far. Screening of inhibitors has been hindered by the lack of a suitable peptide substrate and, consequently, a convenient activity assay. Here we describe the first fluorescence resonance energy transfer (FRET) substrate for ADAMTS-7, ATS7FP7. ATS7FP7 was used to measure inhibition constants for the endogenous ADAMTS-7 inhibitor, TIMP-4, as well as two hydroxamate-based zinc chelating inhibitors. These inhibition constants match well with IC50 values obtained with our SDS-PAGE assay that uses the N-terminal fragment of latent TGF-β-binding protein 4 (LTBP4S-A) as a substrate. Our novel fluorogenic substrate ATS7FP7 is suitable for high throughput screening of ADAMTS-7 inhibitors, thus accelerating translational studies aiming at inhibition of ADAMTS-7 as a novel treatment for cardiovascular diseases such as atherosclerosis and CAD.
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Volume 36
Issue number 1
Pages 2160-2169
Available online
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Publisher website (DOI) 10.1080/14756366.2021.1983808
Europe PubMed Central 34587841
Pubmed 34587841
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