Large-scale expression and purification of a G-protein-coupled receptor for structure determination -- an overview
Reinhard Grisshammer1, Jim F White, Loc B Trinh
1Laboratory of Molecular Biology of the National Institute of Diabetes and Digestive and Kidney Diseases, Department of Health and Human Services, National Institutes of Health, Bethesda, Maryland 20892, USA. rkgriss@helix.nih.gov
Abstract:
Structure determination of G-protein-coupled receptors and other applications, such as nuclear magnetic resonance studies, require milligram quantities of purified, functional receptor protein on a regular basis. We present an overview on expression and purification studies with a receptor for neurotensin. Functional expression in Escherichia coli and an automated two-column purification routine allow ongoing crystallization experiments and studies on receptor-bound ligands.
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