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A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence (FUEL)
Published on: May 23, 2014
Probing biocatalytic transformations with luminescent DNA/silver nanoclusters
Xiaoqing Liu1, Fuan Wang, Angelica Niazov-Elkan
1Institute of Chemistry, Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Nano Letters
|December 21, 2012
Summary
DNA-stabilized silver nanoclusters (AgNCs) offer sensitive fluorescent detection of enzyme activity. These AgNCs effectively probe hydrogen peroxide-generating enzymes and quinone products, enabling ultrasensitive enzyme assays.
Area of Science:
- Biochemistry
- Nanotechnology
- Analytical Chemistry
Background:
- DNA-stabilized silver nanoclusters (AgNCs) are valuable fluorescent labels.
- Hydrogen peroxide (H2O2) and quinones effectively quench AgNC fluorescence.
- This quenching property can be leveraged for enzyme activity detection.
Purpose of the Study:
- To demonstrate the use of AgNCs for probing enzyme activities and their substrates.
- To showcase the detection of H2O2-generating oxidases and quinone-producing enzymes.
- To implement AgNCs for sensitive detection of bienzyme catalytic cascades.
Main Methods:
- Utilized DNA-stabilized AgNCs as fluorescent probes.
- Exploited the fluorescence quenching effect of H2O2 and quinones.
- Applied AgNCs to monitor glucose oxidase, tyrosinase, and bienzyme cascades.
Main Results:
- Successfully probed glucose oxidase activity via H2O2 formation.
- Detected tyrosinase activity by monitoring quinone production from tyrosine, dopamine, or tyramine.
- Achieved ultrasensitive detection of alkaline phosphatase (5 × 10(-5) units/mL) in a bienzyme cascade.
Conclusions:
- DNA-stabilized AgNCs provide a sensitive platform for enzyme activity detection.
- The fluorescence quenching mechanism allows for the probing of various enzymatic reactions.
- AgNCs enable ultrasensitive detection of enzymes and their products in complex biological systems.
