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Medical Research Council

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Automated Protocols for Macromolecular Crystallization at the MRC Laboratory of Molecular Biology

¹Laboratory of Molecular Biology, Medical Research Council

Automated systems and protocols for the routine preparation of a large number of screens and nanoliter crystallization droplets for vapor diffusion experiments are described and discussed.

Biochemistry

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

Filip Pamula^1,2, Jonas Mühle¹, Alain Blanc³, Rony Nehmé⁴, Patricia C. Edwards⁴, Christopher G. Tate⁴, Ching-Ju Tsai¹

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

This report describes screening of different detergents for preparing the visual GPCR, rhodopsin, and its complex with mini-G_o. 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.