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Nanyang Technological University

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

5.3K Views

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10:30 min

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December 27th, 2013

DOI :

10.3791/50908-v

December 27th, 2013

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Dejan Caglič¹, Kristin M. Bompiani¹, Michelle C. Krutein¹, Petr Čapek¹, Tobin J. Dickerson¹^,²

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

Summary

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.

Explore More Videos

Botulinum Neurotoxin

High throughput Screening

Neurotransmitter Release

Therapeutic Target

Chapters in this video

0:05

Title

1:26

Preparation of Compound Dilution Series

2:55

Plate Preparation and Spotting

4:44

Substrate Addition and Fluorescence Measurement

5:32

Automated Operation for High-throughput Screening

7:30

Results: Analyzing the FRET-based Botulinum Neurotoxin/A Light Chain Assay Kinetic Data

9:39

Conclusion

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