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
James B. Duke Professor of Medicine
The focus of work in this laboratory is on the elucidation of the molecular properties
and regulatory mechanisms controlling the function of G protein-coupled receptors.
As model systems we utilize the so called adrenergic receptors for adrenaline and
related molecules. The goal is to learn the general principles of signal transduction
from the outside to the inside of the cell which are involved in systems as diverse
as sensory perception, neuro- transmitter and hormonal signaling. Stud
Articles written by Duke faculty are made available through the campus open access policy. For more information see: Duke Open Access Policy
Rights for Collection: Scholarly Articles
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