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In vitro Investigation of the MexAB Efflux Pump From Pseudomonas aeruginosa
JoVE Journal
Biology
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JoVE Journal Biology
In vitro Investigation of the MexAB Efflux Pump From Pseudomonas aeruginosa
DOI:

13:40 min

February 17, 2014

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Chapters

  • 00:05Title
  • 03:53Protein Production and Purification
  • 05:24Preparation of Proteoliposomes
  • 08:35Assessment of the Reconstitution Efficacy on Sucrose Gradient
  • 09:37Fluorescence Staining
  • 10:28Monitoring Pyranine Fluorescence as a Function of Time While a Proton Gradient is Generated
  • 12:40Conclusion

Summary

Automatic Translation

We present a protocol for the in vitro investigation of efflux pumps from Pseudomonas aeruginosa. This protocol allows for the generation of a robust, reversible, and tunable proton gradient within the liposome membrane and hence should be adaptable to any membrane protein energized by the protomotive force.

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