Purification, crystallization and preliminary X-ray diffraction studies of a complex between G protein-coupled receptor kinase 2 and Gbeta1gamma2.
Abstract
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.
Type
Journal articleSubject
AnimalsCattle
Cell Line
Crystallization
Cyclic AMP-Dependent Protein Kinases
Heterotrimeric GTP-Binding Proteins
Protein Subunits
Recombinant Proteins
Spodoptera
X-Ray Diffraction
beta-Adrenergic Receptor Kinases
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Show full item recordScholars@Duke
Robert J. Lefkowitz
The Chancellor's Distinguished Professor of Medicine
Dr. Lefkowitz’s memoir, A Funny Thing Happened on the Way to Stockholm, recounts his
early career as a cardiologist and his transition to biochemistry, which led to his
Nobel Prize win.
Robert J. Lefkowitz, M.D. is James B. Duke Professor of Medicine and Professor of
Biochemistry and Chemistry at the Duke University Medical Center. He has been an Investigator
of the
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