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JoVE Journal
Immunology and Infection

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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
 
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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking

Article DOI: 10.3791/51217-v 11:33 min December 17th, 2013
December 17th, 2013

Kapitel

Summary

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Click here for the English version.

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.

Tags

Assembly Secreted Bacterial Virulence Factor Site-specific Crosslinking Protein-protein Interactions Amber Suppression Technology Amber Mutation E. Coli Amber Suppressor TRNA Amino Acyl-tRNA Synthetase Methanococcus Jannaschii Photo Activatable Amino Acid Analog P-benzoylphenylalanine (Bpa) Ultraviolet Light Covalent Link Proteins Photocrosslinking Pulse-chase Labeling Immunoprecipitation Bacterial Virulence Factor Assembly Outer Membrane Extracellular Space Prokaryotic Cells Eukaryotic Cells
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