Purification, crystallization and preliminary X-ray diffraction studies of a complex between G protein-coupled receptor kinase 2 and Gbeta1gamma2.
G 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.
Lodowski
David T
DT
Institute for Cellular and Molecular Biology, Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX, USA.
Barnhill
Jennifer F
JF
Pitcher
Julie A
JA
Capel
W Darrell
WD
Lefkowitz
Robert J
RJ
Tesmer
John J G
JJ
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003460:Crystallization / Q000379:methods; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry* / Q000302:isolation & purification*; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D018411:Spodoptera; D014961:X-Ray Diffraction; D051552:beta-Adrenergic Receptor Kinases
D021122:Protein Subunits; D011994:Recombinant Proteins; D017868:Cyclic AMP-Dependent Protein Kinases; D051552:beta-Adrenergic Receptor Kinases; D020962:Heterotrimeric GTP-Binding Proteins