The amino-terminal domain of G-protein-coupled receptor kinase 2
is a regulatory Gbeta gamma binding site
T. Eichmann, K. Lorenz, M. Hoffmann, J. Brockmann, C. Krasel, M. Lohse, and U. Quitterer. J Biol Chem, 278 (10):
8052-7(March 2003)Eichmann, Tanja Lorenz, Kristina Hoffmann, Michaela Brockmann, Jorg
Krasel, Cornelius Lohse, Martin J Quitterer, Ursula Research Support,
Non-U.S. Gov't United States The Journal of biological chemistry
J Biol Chem. 2003 Mar 7;278(10):8052-7. Epub 2002 Dec 16..
Abstract
G-protein-coupled receptor kinase 2 (GRK2) is activated by free Gbetagamma
subunits. A Gbetagamma binding site of GRK2 is localized in the carboxyl-terminal
pleckstrin homology domain. This Gbetagamma binding site of GRK2
also regulates Gbetagamma-stimulated signaling by sequestering free
Gbetagamma subunits. We report here that truncation of the carboxyl-terminal
Gbetagamma binding site of GRK2 did not abolish the Gbetagamma regulatory
activity of GRK2 as determined by the inhibition of a Gbetagamma-stimulated
increase in inositol phosphates in cells. This finding suggested
the presence of a second Gbetagamma binding site in GRK2. And indeed,
the amino terminus of GRK2 (GRK2(1-185)) inhibited a Gbetagamma-stimulated
inositol phosphate signal in cells, purified GRK2(1-185) suppressed
the Gbetagamma-stimulated phosphorylation of rhodopsin, and GRK2(1-185)
bound directly to purified Gbetagamma subunits. The amino-terminal
Gbetagamma regulatory site does not overlap with the RGS domain of
GRK-2 because GRK2(1-53) with truncated RGS domain inhibited Gbetagamma-mediated
signaling with similar potency and efficacy as did GRK2(1-185). In
addition to the Gbetagamma regulatory activity, the amino-terminal
Gbetagamma binding site of GRK2 affects the kinase activity of GRK2
because antibodies specifically cross-reacting with the amino terminus
of GRK2 suppressed the GRK2-dependent phosphorylation of rhodopsin.
The antibody-mediated inhibition was released by purified Gbetagamma
subunits, strongly suggesting that Gbetagamma binding to the amino
terminus of GRK2 enhances the kinase activity toward rhodopsin. Thus,
the amino-terminal domain of GRK2 is a previously unrecognized Gbetagamma
binding site that regulates GRK2-mediated receptor phosphorylation
and inhibits Gbetagamma-stimulated signaling.
Eichmann, Tanja Lorenz, Kristina Hoffmann, Michaela Brockmann, Jorg
Krasel, Cornelius Lohse, Martin J Quitterer, Ursula Research Support,
Non-U.S. Gov't United States The Journal of biological chemistry
J Biol Chem. 2003 Mar 7;278(10):8052-7. Epub 2002 Dec 16.
%0 Journal Article
%1 Eichmann2003
%A Eichmann, T.
%A Lorenz, K.
%A Hoffmann, M.
%A Brockmann, J.
%A Krasel, C.
%A Lohse, M. J.
%A Quitterer, U.
%D 2003
%J J Biol Chem
%K AMP-Dependent Binding Cell Cyclic GTP-Binding Humans Kinases Kinases/chemistry/genetics/*metabolism Line Protein Proteins/chemistry/genetics/metabolism Receptor Recombinant Signal Sites Transduction Transfection beta-Adrenergic Proteins/metabolism
%N 10
%P 8052-7
%T The amino-terminal domain of G-protein-coupled receptor kinase 2
is a regulatory Gbeta gamma binding site
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12486133
%V 278
%X G-protein-coupled receptor kinase 2 (GRK2) is activated by free Gbetagamma
subunits. A Gbetagamma binding site of GRK2 is localized in the carboxyl-terminal
pleckstrin homology domain. This Gbetagamma binding site of GRK2
also regulates Gbetagamma-stimulated signaling by sequestering free
Gbetagamma subunits. We report here that truncation of the carboxyl-terminal
Gbetagamma binding site of GRK2 did not abolish the Gbetagamma regulatory
activity of GRK2 as determined by the inhibition of a Gbetagamma-stimulated
increase in inositol phosphates in cells. This finding suggested
the presence of a second Gbetagamma binding site in GRK2. And indeed,
the amino terminus of GRK2 (GRK2(1-185)) inhibited a Gbetagamma-stimulated
inositol phosphate signal in cells, purified GRK2(1-185) suppressed
the Gbetagamma-stimulated phosphorylation of rhodopsin, and GRK2(1-185)
bound directly to purified Gbetagamma subunits. The amino-terminal
Gbetagamma regulatory site does not overlap with the RGS domain of
GRK-2 because GRK2(1-53) with truncated RGS domain inhibited Gbetagamma-mediated
signaling with similar potency and efficacy as did GRK2(1-185). In
addition to the Gbetagamma regulatory activity, the amino-terminal
Gbetagamma binding site of GRK2 affects the kinase activity of GRK2
because antibodies specifically cross-reacting with the amino terminus
of GRK2 suppressed the GRK2-dependent phosphorylation of rhodopsin.
The antibody-mediated inhibition was released by purified Gbetagamma
subunits, strongly suggesting that Gbetagamma binding to the amino
terminus of GRK2 enhances the kinase activity toward rhodopsin. Thus,
the amino-terminal domain of GRK2 is a previously unrecognized Gbetagamma
binding site that regulates GRK2-mediated receptor phosphorylation
and inhibits Gbetagamma-stimulated signaling.
@article{Eichmann2003,
abstract = {G-protein-coupled receptor kinase 2 (GRK2) is activated by free Gbetagamma
subunits. A Gbetagamma binding site of GRK2 is localized in the carboxyl-terminal
pleckstrin homology domain. This Gbetagamma binding site of GRK2
also regulates Gbetagamma-stimulated signaling by sequestering free
Gbetagamma subunits. We report here that truncation of the carboxyl-terminal
Gbetagamma binding site of GRK2 did not abolish the Gbetagamma regulatory
activity of GRK2 as determined by the inhibition of a Gbetagamma-stimulated
increase in inositol phosphates in cells. This finding suggested
the presence of a second Gbetagamma binding site in GRK2. And indeed,
the amino terminus of GRK2 (GRK2(1-185)) inhibited a Gbetagamma-stimulated
inositol phosphate signal in cells, purified GRK2(1-185) suppressed
the Gbetagamma-stimulated phosphorylation of rhodopsin, and GRK2(1-185)
bound directly to purified Gbetagamma subunits. The amino-terminal
Gbetagamma regulatory site does not overlap with the RGS domain of
GRK-2 because GRK2(1-53) with truncated RGS domain inhibited Gbetagamma-mediated
signaling with similar potency and efficacy as did GRK2(1-185). In
addition to the Gbetagamma regulatory activity, the amino-terminal
Gbetagamma binding site of GRK2 affects the kinase activity of GRK2
because antibodies specifically cross-reacting with the amino terminus
of GRK2 suppressed the GRK2-dependent phosphorylation of rhodopsin.
The antibody-mediated inhibition was released by purified Gbetagamma
subunits, strongly suggesting that Gbetagamma binding to the amino
terminus of GRK2 enhances the kinase activity toward rhodopsin. Thus,
the amino-terminal domain of GRK2 is a previously unrecognized Gbetagamma
binding site that regulates GRK2-mediated receptor phosphorylation
and inhibits Gbetagamma-stimulated signaling.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Eichmann, T. and Lorenz, K. and Hoffmann, M. and Brockmann, J. and Krasel, C. and Lohse, M. J. and Quitterer, U.},
biburl = {https://www.bibsonomy.org/bibtex/246fdfe4b43d4e43908149929d339d6a6/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {c3097e398db3284a589e29b6df857400},
intrahash = {46fdfe4b43d4e43908149929d339d6a6},
issn = {0021-9258 (Print) 0021-9258 (Linking)},
journal = {J Biol Chem},
keywords = {AMP-Dependent Binding Cell Cyclic GTP-Binding Humans Kinases Kinases/chemistry/genetics/*metabolism Line Protein Proteins/chemistry/genetics/metabolism Receptor Recombinant Signal Sites Transduction Transfection beta-Adrenergic Proteins/metabolism},
month = {Mar 7},
note = {Eichmann, Tanja Lorenz, Kristina Hoffmann, Michaela Brockmann, Jorg
Krasel, Cornelius Lohse, Martin J Quitterer, Ursula Research Support,
Non-U.S. Gov't United States The Journal of biological chemistry
J Biol Chem. 2003 Mar 7;278(10):8052-7. Epub 2002 Dec 16.},
number = 10,
pages = {8052-7},
shorttitle = {The amino-terminal domain of G-protein-coupled receptor kinase 2 is
a regulatory Gbeta gamma binding site},
timestamp = {2010-12-14T18:21:42.000+0100},
title = {The amino-terminal domain of G-protein-coupled receptor kinase 2
is a regulatory Gbeta gamma binding site},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12486133},
volume = 278,
year = 2003
}