Large-scale expression and purification of a G-protein-coupled receptor for structure determination -- an overview.
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.
Grisshammer
Reinhard
R
Laboratory 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
White
Jim F
JF
Trinh
Loc B
LB
Shiloach
Joseph
J
D003460:Crystallization / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D011487:Protein Conformation; D040901:Proteomics / Q000379:methods*; D018028:Receptors, Neurotensin / Q000302:isolation & purification* / Q000378:metabolism*
D018028:Receptors, Neurotensin