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

Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization

6,696 Views

•

09:19 min

•

March 16, 2020

DOI:

10.3791/60747-v

DOI:
10.3791/60747-v

•

09:19 min
March 16, 2020

•

2 Views
, , , , , ,
1Laboratory of Biomolecular Research,Paul Scherrer Institute, 2Department of Biology,ETH Zürich, 3Center for Radiopharmaceutical Sciences,Paul Scherrer Institute, 4Laboratory of Molecular Biology,Medical Research Council

Summary

Automatically generated

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.

Explore More Videos

GPCRMini-G ProteinSignal TransductionMembrane Protein ComplexDetergent ScreeningCrystallizationHEK293 CellsAffinity Purification9-cis RetinalBuffer Exchange

Read Article

Cite this Article

Pamula, F., Mühle, J., Blanc, A., Nehmé, R., Edwards, P. C., Tate, C. G., Tsai, C. Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization. J. Vis. Exp. (157), e60747, doi:10.3791/60747 (2020).

Download .ris file

Copy

Share Video

.

Copy