A Fluorogenic Peptide Cleavage Assay to Screen for Proteolytic Activity: Applications for coronavirus spike protein activation

33.0K Views

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07:53 min

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January 9th, 2019

DOI :

10.3791/58892-v

January 9th, 2019

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Javier A. Jaimes*¹^,², Jean K. Millet*¹^,³, Monty E. Goldstein¹^,⁴, Gary R. Whittaker¹, Marco R. Straus¹

¹Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, ²Department of Microbiology, College of Agricultural and Life Sciences, Cornell University, ³Virologie et Immunologie Moléculaires, Domaine de Vilvert, INRA, ⁴Department of Cell Biology and Molecular Genetics, University of Maryland

Summary

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Fluorogenic Peptide Cleavage Assay

Proteolytic Activity

Coronavirus Spike Protein

Protease Recognition Site

Fluorescence Resonance Energy Transfer

Fluorescence Measurement

Chapters in this video

0:04

Title

0:31

Designing and Preparing the Peptides

2:06

Preparing the Fluorescence Plate Reader

3:11

Preparing the Assay

4:30

Data Analysis

5:37

Results: Mutations in the Cleavage Site Alter Activity of Furin and Cleavage Efficiency

7:26

Conclusion

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