Purification, crystallization and preliminary X-ray diffraction studies of a complex between G protein-coupled receptor kinase 2 and Gbeta1gamma2.

Lodowski, David TBarnhill, Jennifer FPitcher, Julie ACapel, W DarrellLefkowitz, Robert JTesmer, John JG2013-09-052003-050907-4449https://hdl.handle.net/10161/7801G protein-coupled receptor kinase 2 (GRK2) phosphorylates activated G protein-coupled receptors (GPCRs), which ultimately leads to their desensitization and/or downregulation. The enzyme is recruited to the plasma membrane via the interaction of its carboxyl-terminal pleckstrin-homology (PH) domain with the beta and gamma subunits of heterotrimeric G proteins (Gbetagamma). An improved purification scheme for GRK2 has been developed, conditions under which GRK2 forms a complex with Gbeta(1)gamma(2) have been determined and the complex has been crystallized in CHAPS detergent micelles. Crystals of the GRK2-Gbetagamma complex belong to space group C2 and have unit-cell parameters a = 187.0, b = 72.1, c = 122.0 A, beta = 115.2 degrees. A complete data set has been collected to 3.2 A resolution with Cu Kalpha radiation.AnimalsCattleCell LineCrystallizationCyclic AMP-Dependent Protein KinasesHeterotrimeric GTP-Binding ProteinsProtein SubunitsRecombinant ProteinsSpodopteraX-Ray Diffractionbeta-Adrenergic Receptor KinasesPurification, crystallization and preliminary X-ray diffraction studies of a complex between G protein-coupled receptor kinase 2 and Gbeta1gamma2.Journal article