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A High-throughput-compatible FRET-based Platform for Identification and Characterization of Botulinum Neurotoxin Light Chain Modulators
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Chemistry
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JoVE Journal Chemistry
A High-throughput-compatible FRET-based Platform for Identification and Characterization of Botulinum Neurotoxin Light Chain Modulators

A High-throughput-compatible FRET-based Platform for Identification and Characterization of Botulinum Neurotoxin Light Chain Modulators

DOI:
10.3791/50908-v

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10:30 min

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December 27, 2013

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, , , ,
1Department of Chemistry,The Scripps Research Institute, 2Worm Institute of Research and Medicine (WIRM),The Scripps Research Institute

Chapters

  • 00:05Title
  • 01:26Preparation of Compound Dilution Series
  • 02:55Plate Preparation and Spotting
  • 04:44Substrate Addition and Fluorescence Measurement
  • 05:32Automated Operation for High-throughput Screening
  • 07:30Results: Analyzing the FRET-based Botulinum Neurotoxin/A Light Chain Assay Kinetic Data
  • 09:39Conclusion

Summary

Automatic Translation

Automatic Translation

The botulinum neurotoxin type A light chain (BoNT/A LC) is a metalloprotease that enters motor neurons, cleaves its substrate SNAP-25, and disrupts neurotransmission, thereby resulting in flaccid paralysis. Utilizing a high-throughput-compatible FRET-based assay, large libraries of small molecules can be screened for their impact on BoNT/A LC enzymatic activity.

Tags

Botulinum NeurotoxinBoNTLight ChainFRETHigh-throughput ScreeningHTSInhibitorAcetylcholineNeurotransmitter ReleaseTherapeutic TargetSmall Molecule

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