Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic
receptor kinase, cAMP-dependent protein kinase, and protein kinase
C occurs via distinct molecular mechanisms
J. Pitcher, M. Lohse, J. Codina, M. Caron, und R. Lefkowitz. Biochemistry, 31 (12):
3193-7(März 1992)Pitcher, J Lohse, M J Codina, J Caron, M G Lefkowitz, R J HL16037/HL/NHLBI
NIH HHS/United States Research Support, U.S. Gov't, P.H.S. United
states Biochemistry Biochemistry. 1992 Mar 31;31(12):3193-7..
Zusammenfassung
Exposure of beta 2-adrenergic receptors (beta 2ARs) to agonists causes
a rapid desensitization of the receptor-stimulated adenylyl cyclase
response. Phosphorylation of the beta 2AR by several distinct kinases
plays an important role in this desensitization phenomenon. In this
study, we have utilized purified hamster lung beta 2AR and stimulatory
guanine nucleotide binding regulatory protein (Gs), reconstituted
in phospholipid vesicles, to investigate the molecular properties
of this desensitization response. Purified hamster beta 2AR was phosphorylated
by cAMP-dependent protein kinase (PKA), protein kinase C (PKC), or
beta AR kinase (beta ARK), and receptor function was determined by
measuring the beta 2AR-agonist-promoted Gs-associated GTPase activity.
At physiological concentrations of Mg2+ (less than 1 mM), receptor
phosphorylation inhibited coupling to Gs by 60% (PKA), 40% (PKC),
and 30% (beta ARK). The desensitizing effect of phosphorylation was,
however, greatly diminished when assays were performed at concentrations
of Mg2+ sufficient to promote receptor-independent activation of
Gs (greater than 5 mM). Addition of retinal arrestin, the light transduction
component involved in the attenuation of rhodopsin function, did
not enhance the uncoupling effect of beta ARK phosphorylation of
beta 2AR when assayed in the presence of 0.3 mM free Mg2+. At concentrations
of Mg2+ ranging between 0.5 and 5.0 mM, however, significant potentiation
of beta ARK-mediated desensitization was observed upon arrestin addition.
At a free Mg2+ concentration of 5 mM, arrestin did not potentiate
the inhibition of receptor function observed on PKA or PKC phosphorylation.
These results suggest that distinct pathways of desensitization exist
for the receptor phosphorylated either by PKA or PKC or alternatively
by beta ARK.
Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic
receptor kinase, cAMP-dependent protein kinase, and protein kinase
C occurs via distinct molecular mechanisms
Pitcher, J Lohse, M J Codina, J Caron, M G Lefkowitz, R J HL16037/HL/NHLBI
NIH HHS/United States Research Support, U.S. Gov't, P.H.S. United
states Biochemistry Biochemistry. 1992 Mar 31;31(12):3193-7.
%0 Journal Article
%1 Pitcher1992
%A Pitcher, J.
%A Lohse, M. J.
%A Codina, J.
%A Caron, M. G.
%A Lefkowitz, R. J.
%D 1992
%J Biochemistry
%K & *Cyclic AMP-Dependent AMP/physiology Adrenergic Animals Antigens/physiology Arrestin C/*pharmacology Cricetinae Cyclic Drug Eye Humans Kinase Kinases Kinases/*pharmacology Lung/chemistry Phosphorylation Protein Proteins/physiology Receptor Retina/chemistry Synergism beta-Adrenergic beta-Agonists/*pharmacology beta/*drug effects/isolation purification
%N 12
%P 3193-7
%T Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic
receptor kinase, cAMP-dependent protein kinase, and protein kinase
C occurs via distinct molecular mechanisms
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1348186
%V 31
%X Exposure of beta 2-adrenergic receptors (beta 2ARs) to agonists causes
a rapid desensitization of the receptor-stimulated adenylyl cyclase
response. Phosphorylation of the beta 2AR by several distinct kinases
plays an important role in this desensitization phenomenon. In this
study, we have utilized purified hamster lung beta 2AR and stimulatory
guanine nucleotide binding regulatory protein (Gs), reconstituted
in phospholipid vesicles, to investigate the molecular properties
of this desensitization response. Purified hamster beta 2AR was phosphorylated
by cAMP-dependent protein kinase (PKA), protein kinase C (PKC), or
beta AR kinase (beta ARK), and receptor function was determined by
measuring the beta 2AR-agonist-promoted Gs-associated GTPase activity.
At physiological concentrations of Mg2+ (less than 1 mM), receptor
phosphorylation inhibited coupling to Gs by 60% (PKA), 40% (PKC),
and 30% (beta ARK). The desensitizing effect of phosphorylation was,
however, greatly diminished when assays were performed at concentrations
of Mg2+ sufficient to promote receptor-independent activation of
Gs (greater than 5 mM). Addition of retinal arrestin, the light transduction
component involved in the attenuation of rhodopsin function, did
not enhance the uncoupling effect of beta ARK phosphorylation of
beta 2AR when assayed in the presence of 0.3 mM free Mg2+. At concentrations
of Mg2+ ranging between 0.5 and 5.0 mM, however, significant potentiation
of beta ARK-mediated desensitization was observed upon arrestin addition.
At a free Mg2+ concentration of 5 mM, arrestin did not potentiate
the inhibition of receptor function observed on PKA or PKC phosphorylation.
These results suggest that distinct pathways of desensitization exist
for the receptor phosphorylated either by PKA or PKC or alternatively
by beta ARK.
@article{Pitcher1992,
abstract = {Exposure of beta 2-adrenergic receptors (beta 2ARs) to agonists causes
a rapid desensitization of the receptor-stimulated adenylyl cyclase
response. Phosphorylation of the beta 2AR by several distinct kinases
plays an important role in this desensitization phenomenon. In this
study, we have utilized purified hamster lung beta 2AR and stimulatory
guanine nucleotide binding regulatory protein (Gs), reconstituted
in phospholipid vesicles, to investigate the molecular properties
of this desensitization response. Purified hamster beta 2AR was phosphorylated
by cAMP-dependent protein kinase (PKA), protein kinase C (PKC), or
beta AR kinase (beta ARK), and receptor function was determined by
measuring the beta 2AR-agonist-promoted Gs-associated GTPase activity.
At physiological concentrations of Mg2+ (less than 1 mM), receptor
phosphorylation inhibited coupling to Gs by 60% (PKA), 40% (PKC),
and 30% (beta ARK). The desensitizing effect of phosphorylation was,
however, greatly diminished when assays were performed at concentrations
of Mg2+ sufficient to promote receptor-independent activation of
Gs (greater than 5 mM). Addition of retinal arrestin, the light transduction
component involved in the attenuation of rhodopsin function, did
not enhance the uncoupling effect of beta ARK phosphorylation of
beta 2AR when assayed in the presence of 0.3 mM free Mg2+. At concentrations
of Mg2+ ranging between 0.5 and 5.0 mM, however, significant potentiation
of beta ARK-mediated desensitization was observed upon arrestin addition.
At a free Mg2+ concentration of 5 mM, arrestin did not potentiate
the inhibition of receptor function observed on PKA or PKC phosphorylation.
These results suggest that distinct pathways of desensitization exist
for the receptor phosphorylated either by PKA or PKC or alternatively
by beta ARK.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Pitcher, J. and Lohse, M. J. and Codina, J. and Caron, M. G. and Lefkowitz, R. J.},
biburl = {https://www.bibsonomy.org/bibtex/2e4afb61d620bd42c6557d89176840b3e/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {c7810a8ef711c3e897eca9a9b5edaba4},
intrahash = {e4afb61d620bd42c6557d89176840b3e},
issn = {0006-2960 (Print) 0006-2960 (Linking)},
journal = {Biochemistry},
keywords = {& *Cyclic AMP-Dependent AMP/physiology Adrenergic Animals Antigens/physiology Arrestin C/*pharmacology Cricetinae Cyclic Drug Eye Humans Kinase Kinases Kinases/*pharmacology Lung/chemistry Phosphorylation Protein Proteins/physiology Receptor Retina/chemistry Synergism beta-Adrenergic beta-Agonists/*pharmacology beta/*drug effects/isolation purification},
month = {Mar 31},
note = {Pitcher, J Lohse, M J Codina, J Caron, M G Lefkowitz, R J HL16037/HL/NHLBI
NIH HHS/United States Research Support, U.S. Gov't, P.H.S. United
states Biochemistry Biochemistry. 1992 Mar 31;31(12):3193-7.},
number = 12,
pages = {3193-7},
shorttitle = {Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic
receptor kinase, cAMP-dependent protein kinase, and protein kinase
C occurs via distinct molecular mechanisms},
timestamp = {2010-12-14T18:22:42.000+0100},
title = {Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic
receptor kinase, cAMP-dependent protein kinase, and protein kinase
C occurs via distinct molecular mechanisms},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1348186},
volume = 31,
year = 1992
}