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Related Experiment Video

Updated: Oct 18, 2025

Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
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Development of a fluorogenic ADAMTS-7 substrate.

Salvatore Santamaria1, Frederic Buemi1, Elisa Nuti2

  • 1Department of Immunology and Inflammation, Imperial College London, London, UK.

Journal of Enzyme Inhibition and Medicinal Chemistry
|September 30, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel FRET substrate, ATS7FP7, to screen for ADAMTS-7 inhibitors. This breakthrough accelerates the development of new treatments for atherosclerosis and coronary artery disease (CAD).

Keywords:
ADAMTS-7ADAMTS7activity assaycoronary artery diseaseinhibitor

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Area of Science:

  • Biochemistry and Molecular Biology
  • Cardiovascular Research
  • Drug Discovery

Background:

  • ADAMTS-7, an extracellular protease, is a potential therapeutic target for atherosclerosis and coronary artery disease (CAD).
  • Development of ADAMTS-7 inhibitors has been limited by the absence of suitable peptide substrates and activity assays.
  • A convenient assay is crucial for high-throughput screening (HTS) of potential ADAMTS-7 inhibitors.

Purpose of the Study:

  • To develop the first fluorescence resonance energy transfer (FRET) substrate for ADAMTS-7.
  • To utilize the novel substrate for measuring inhibition constants of known inhibitors.
  • To enable high-throughput screening for novel ADAMTS-7 inhibitors to treat cardiovascular diseases.

Main Methods:

  • Development and characterization of a novel FRET peptide substrate (ATS7FP7) for ADAMTS-7.
  • Measurement of inhibition constants (Ki) for TIMP-4 and hydroxamate-based inhibitors using ATS7FP7.
  • Comparison of FRET assay results with IC50 values from an established SDS-PAGE assay using LTBP4S-A substrate.

Main Results:

  • The novel FRET substrate, ATS7FP7, was successfully developed for ADAMTS-7 activity.
  • Inhibition constants measured with ATS7FP7 correlated well with IC50 values from the SDS-PAGE assay.
  • ATS7FP7 demonstrated suitability for measuring inhibition of ADAMTS-7 by endogenous and synthetic inhibitors.

Conclusions:

  • The fluorogenic FRET substrate ATS7FP7 is a valuable tool for quantifying ADAMTS-7 activity and inhibition.
  • ATS7FP7 facilitates high-throughput screening, paving the way for discovering novel ADAMTS-7 inhibitors.
  • This advancement is expected to accelerate translational research for new cardiovascular disease treatments targeting ADAMTS-7.