Show simple item record

Monoclonal antibodies reveal receptor specificity among G-protein-coupled receptor kinases.

dc.contributor.author Diversé-Pierluissi, M
dc.contributor.author Drazner, Mark H
dc.contributor.author Dyer, SL
dc.contributor.author Freedman, NJ
dc.contributor.author Lefkowitz, Robert J
dc.contributor.author Oppermann, M
dc.contributor.author Peppel, Karsten C
dc.coverage.spatial United States
dc.date.accessioned 2013-09-10T15:00:50Z
dc.date.issued 1996-07-23
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/8755530
dc.identifier.issn 0027-8424
dc.identifier.uri http://hdl.handle.net/10161/7835
dc.description.abstract Guanine nucleotide-binding regulatory protein (G protein)-coupled receptor kinases (GRKs) constitute a family of serine/threonine kinases that play a major role in the agonist-induced phosphorylation and desensitization of G-protein-coupled receptors. Herein we describe the generation of monoclonal antibodies (mAbs) that specifically react with GRK2 and GRK3 or with GRK4, GRK5, and GRK6. They are used in several different receptor systems to identify the kinases that are responsible for receptor phosphorylation and desensitization. The ability of these reagents to inhibit GRK- mediated receptor phosphorylation is demonstrated in permeabilized 293 cells that overexpress individual GRKs and the type 1A angiotensin II receptor. We also use this approach to identify the endogenous GRKs that are responsible for the agonist-induced phosphorylation of epitope-tagged beta2- adrenergic receptors (beta2ARs) overexpressed in rabbit ventricular myocytes that are infected with a recombinant adenovirus. In these myocytes, anti-GRK2/3 mAbs inhibit isoproterenol-induced receptor phosphorylation by 77%, while GRK4-6-specific mAbs have no effect. Consistent with the operation of a betaAR kinase-mediated mechanism, GRK2 is identified by immunoblot analysis as well as in a functional assay as the predominant GRK expressed in these cells. Microinjection of GRK2/3-specific mAbs into chicken sensory neurons, which have been shown to express a GRK3-like protein, abolishes desensitization of the alpha2AR-mediated calcium current inhibition. The intracellular inhibition of endogenous GRKs by mAbs represents a novel approach to the study of receptor specificities among GRKs that should be widely applicable to many G-protein-coupled receptors.
dc.language eng
dc.relation.ispartof Proc Natl Acad Sci U S A
dc.subject Angiotensin II
dc.subject Animals
dc.subject Antibodies, Monoclonal
dc.subject Antibody Specificity
dc.subject Cell Line
dc.subject Cells, Cultured
dc.subject Chickens
dc.subject Cyclic AMP-Dependent Protein Kinases
dc.subject G-Protein-Coupled Receptor Kinase 3
dc.subject G-Protein-Coupled Receptor Kinases
dc.subject GTP-Binding Proteins
dc.subject Heart
dc.subject Heart Ventricles
dc.subject Humans
dc.subject Immunoblotting
dc.subject Isoproterenol
dc.subject Kinetics
dc.subject Membrane Proteins
dc.subject Myocardium
dc.subject Neurons, Afferent
dc.subject Phosphorylation
dc.subject Protein-Serine-Threonine Kinases
dc.subject Rabbits
dc.subject Receptor Protein-Tyrosine Kinases
dc.subject Receptors, Cell Surface
dc.subject Receptors, G-Protein-Coupled
dc.subject Transfection
dc.subject beta-Adrenergic Receptor Kinases
dc.title Monoclonal antibodies reveal receptor specificity among G-protein-coupled receptor kinases.
dc.type Journal article
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/8755530
pubs.begin-page 7649
pubs.end-page 7654
pubs.issue 15
pubs.organisational-group Basic Science Departments
pubs.organisational-group Biochemistry
pubs.organisational-group Cell Biology
pubs.organisational-group Chemistry
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Duke
pubs.organisational-group Duke Cancer Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Medicine
pubs.organisational-group Medicine, Cardiology
pubs.organisational-group Pathology
pubs.organisational-group School of Medicine
pubs.organisational-group Trinity College of Arts & Sciences
pubs.publication-status Published
pubs.volume 93


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record