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Related Experiment Videos

Novel fluorogenic substrates for imaging beta-lactamase gene expression.

Wenzhong Gao1, Bengang Xing, Roger Y Tsien

  • 1Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, University of California, Los Angeles, California 90095-1770, USA.

Journal of the American Chemical Society
|October 14, 2005
PubMed
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New fluorogenic substrates offer a 153-fold fluorescence enhancement upon beta-lactamase hydrolysis. This breakthrough enables sensitive detection of beta-lactamase in cell lysates and imaging in living cells for diagnostics and biosensing.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Beta-lactamase is an enzyme with significant applications in diagnostics and biosensing.
  • Existing fluorogenic substrates for beta-lactamase detection have limitations in sensitivity and signal enhancement.

Purpose of the Study:

  • To develop a novel class of small nonfluorescent fluorogenic substrates for beta-lactamase detection.
  • To achieve significant fluorescence enhancement upon enzymatic hydrolysis.
  • To enable sensitive detection and imaging applications of beta-lactamase.

Main Methods:

  • Synthesis of a new class of nonfluorescent fluorogenic substrates.
  • Hydrolysis of substrates by beta-lactamase enzyme.
  • Measurement of fluorescence intensity enhancement.

Related Experiment Videos

  • Detection of beta-lactamase in cell lysates.
  • Imaging of beta-lactamase expression in living cells using a red fluorescence derivative.

    • Main Results:

    • The new substrates exhibit up to 153-fold enhancement in fluorescence intensity after beta-lactamase hydrolysis.
    • Detection of beta-lactamase at concentrations as low as 500 fM in cell lysates.
    • Successful imaging of beta-lactamase expression in living cells.

    Conclusions:

    • The developed fluorogenic substrates represent a significant advancement for beta-lactamase detection.
    • These substrates offer high sensitivity and signal amplification, suitable for clinical diagnostics.
    • The utility of beta-lactamase as a biosensor is greatly facilitated by these novel substrates.