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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
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Chapitres
- 00:05Titre
- 01:55Culture Preparation
- 02:59Bpa Incorporation and Preparation for Photocrosslinking
- 04:51Pulse-chase Labeling and UV Irradiation
- 07:33Immunoprecipitation and Crosslinking Product Detection
- 08:27Results: Interaction of the Autotransporter EspP wirh Molecular Chaperones and Components of the Bam Complex
- 10:54Conclusion
This article illustrates the use of pulse-chase radio labeling in combination with site-specific photocrosslinking to monitor interactions between a protein of interest and other factors in E. coli. Unlike traditional chemical cross-linking methods, this approach generates high resolution “snapshots” of an ordered assembly pathway in a living cell.
