Development of a fluorogenic ADAMTS-7 substrate
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
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.
Date Issued
2021-09-30
Date Acceptance
2021-09-19
Citation
Journal of Enzyme Inhibition and Medicinal Chemistry, 2021, 36 (1)
ISSN
1475-6366
Publisher
Taylor & Francis Open Access
Journal / Book Title
Journal of Enzyme Inhibition and Medicinal Chemistry
Volume
36
Issue
1
Copyright Statement
© 2021 The Author(s). Published by InformaUK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Sponsor
British Heart Foundation
British Heart Foundation
Grant Number
FS/IBSRF/20/25032
PG/18/19/33584
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Chemistry, Medicinal
Pharmacology & Pharmacy
ADAMTS-7
ADAMTS7
activity assay
inhibitor
coronary artery disease
ARYLSULFONAMIDE INHIBITORS
CLEAVAGE
DOMAIN
AGGRECAN
BINDING
SITE
ADAMTS-7
ADAMTS7
activity assay
coronary artery disease
inhibitor
Medicinal & Biomolecular Chemistry
0304 Medicinal and Biomolecular Chemistry
0601 Biochemistry and Cell Biology
Publication Status
Published
Date Publish Online
2021-09-30