JoVE Journal

Chemistry

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Nanyang Technological University

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

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.

Chapters in this video

0:05

Title

1:26

Preparation of Compound Dilution Series

4:44

Substrate Addition and Fluorescence Measurement

9:39

Conclusion

2:55

Plate Preparation and Spotting

5:32

Automated Operation for High-throughput Screening

7:30

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

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