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Sensing caspase 3 activity with quantum dot-fluorescent protein assemblies.

Kelly Boeneman1, Bing C Mei, Allison M Dennis

  • 1Center for Bio/Molecular Science & Engineering, U.S. Naval Research Laboratory, Washington, DC 20375, USA.

Journal of the American Chemical Society
|February 27, 2009
PubMed
Summary

We developed a novel quantum dot (QD) sensor for detecting caspase 3 enzyme activity. This biosensor uses a fluorescent protein linked to QDs, enabling specific and quantitative monitoring of protease function.

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Area of Science:

  • Biotechnology
  • Biochemistry
  • Materials Science

Background:

  • Protease activity, particularly of caspase 3, is crucial in cellular processes and disease.
  • Existing methods for monitoring protease activity can be complex or lack specificity.

Purpose of the Study:

  • To develop a sensitive and specific biosensor for quantifying caspase 3 proteolytic activity.
  • To utilize a hybrid fluorescent protein-semiconductor quantum dot (QD) system for protease detection.

Main Methods:

  • Engineered mCherry fluorescent protein with a caspase 3 cleavage site.
  • Self-assembled QD-mCherry conjugates via metal-affinity coordination.
  • Utilized fluorescence resonance energy transfer (FRET) for signal transduction.

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Last Updated: Jun 25, 2026

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila
13:21

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila

Published on: November 27, 2016

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry
05:29

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Published on: March 24, 2023

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation
08:47

Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation

Published on: March 5, 2018

Main Results:

  • Demonstrated specific cleavage of the mCherry linker by caspase 3.
  • Observed altered FRET efficiency upon enzymatic cleavage, enabling quantitative monitoring.
  • Achieved ratiometric self-assembly of the sensor components.

Conclusions:

  • The QD-mCherry sensor provides a robust platform for specific caspase 3 activity monitoring.
  • The system offers advantages like bacterial expression, facile assembly, and potential for modification to target other proteases.