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Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
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Ultrasensitive small molecule fluorogenic probe for human heparanase.

Jun Liu1,2, Kelton A Schleyer1,2, Tyrel L Bryan1

  • 1Department of Chemistry and Chemical Biology, University of New Mexico Albuquerque NM 87131 USA.

Chemical Science
|June 24, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel fluorogenic probe for detecting heparanase (HPA) activity. This ultrasensitive tool enables real-time monitoring and inhibitor screening for HPA, an enzyme linked to cancer and inflammation.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Heparanase (HPA) is an enzyme crucial for extracellular matrix (ECM) remodeling.
  • Elevated HPA levels are associated with diseases like cancer and inflammation.
  • Accurate detection of HPA activity is vital for understanding disease mechanisms and developing targeted therapies, but structurally defined probes are lacking.

Purpose of the Study:

  • To develop the first ultrasensitive, structurally defined fluorogenic small-molecule probe for detecting heparanase enzymatic activity.
  • To enable real-time, one-step detection of HPA activity with high sensitivity.
  • To facilitate the discovery of novel molecular therapeutics targeting heparanase.

Main Methods:

  • Development of a novel fluorogenic small-molecule probe by tuning substrate electronic effects.
  • Characterization of probe performance, including fluorescence response and detection limits.
  • Application of the probe in a high-throughput screening assay for heparanase inhibitors.

Main Results:

  • The developed probe exhibits a 756-fold fluorescence turn-on response in the presence of human heparanase.
  • Achieved a picomolar detection limit for HPA activity, enabling ultrasensitive detection.
  • Demonstrated the probe's high sensitivity and robustness in a high-throughput screening assay.

Conclusions:

  • The novel fluorogenic probe represents a significant advancement for detecting heparanase activity.
  • This tool allows for real-time monitoring and sensitive quantification of HPA.
  • The probe is valuable for advancing research in HPA-related diseases and for high-throughput screening of heparanase inhibitors.