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Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization
Journal JoVE
Biochimie
This content is Free Access.
Journal JoVE Biochimie
Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization
DOI:

09:19 min

March 16, 2020

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Chapitres

  • 00:04Introduction
  • 00:45Cell Membrane Solubilization and Protein Extraction
  • 03:53UV-VIS Spectroscopy
  • 04:33Automated Size-Exclusion Chromatography of Rhodopsin and Rhodopsin Mini-Go Complex
  • 06:04Deglycosylation
  • 06:44Results: UV-VIS Spectroscopy, SEC, and SDS-PAGE
  • 08:34Conclusion

Summary

Traduction automatique

This report describes screening of different detergents for preparing the visual GPCR, rhodopsin, and its complex with mini-Go. Biochemical methods characterizing the quality of the complex at different stages during purification are demonstrated. This protocol can be generalized to other membrane protein complexes for their future structural studies.

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