Comparative rates of desensitization of beta-adrenergic receptors
by the beta-adrenergic receptor kinase and the cyclic AMP-dependent
protein kinase
N. Roth, P. Campbell, M. Caron, R. Lefkowitz, and M. Lohse. Proc Natl Acad Sci U S A, 88 (14):
6201-4(July 1991)Roth, N S Campbell, P T Caron, M G Lefkowitz, R J Lohse, M J HL16037/HL/NHLBI
NIH HHS/United States Comparative Study Research Support, U.S. Gov't,
P.H.S. United states Proceedings of the National Academy of Sciences
of the United States of America Proc Natl Acad Sci U S A. 1991 Jul
15;88(14):6201-4..
Abstract
Three separate processes may contribute to rapid beta-adrenergic receptor
desensitization: functional uncoupling from the stimulatory guanine
nucleotide-binding protein Gs, mediated by phosphorylation of the
receptors by two distinct kinases, the specific beta-adrenergic receptor
kinase (beta ARK) and the cyclic AMP-dependent protein kinase A (PKA),
as well as a spatial uncoupling via sequestration of the receptors
away from the cell surface. To evaluate the relative importance and
potential role of the various processes in different physiological
situations, a kinetic analysis of these three mechanisms was performed
in permeabilized A431 epidermoid carcinoma cells. To allow a separate
analysis of each mechanism, inhibitors of the various desensitization
mechanisms were used: heparin to inhibit beta ARK, the PKA inhibitor
peptide PKI to inhibit PKA, and concanavalin A treatment to prevent
sequestration. Isoproterenol-induced phosphorylation of beta 2 receptors
in these cells by beta ARK occurred with a t1/2 of less than 20 sec,
whereas phosphorylation by PKA had a t1/2 of about 2 min. Similarly,
beta ARK-mediated desensitization of the receptors proceeded with
a t1/2 of less than 15 sec, and PKA-mediated desensitization with
a t1/2 of about 3.5 min. Maximal desensitization mediated by the
two kinases corresponded to a reduction of the signal-transduction
capacity of the receptor/adenylyl cyclase system by about 60% in
the case of beta ARK and by about 40% in the case of PKA. Receptor
sequestration was much slower (t1/2 of about 10 min) and involved
no more than 30% of the cell surface receptors. It is concluded that
beta ARK-mediated phosphorylation is the most rapid and quantitatively
most important factor contributing to the rapid desensitization.
This rapidity of the beta ARK-mediated mechanism makes it particularly
well suited to regulate beta-adrenergic receptor function in rapidly
changing environments such as the synaptic cleft.
Roth, N S Campbell, P T Caron, M G Lefkowitz, R J Lohse, M J HL16037/HL/NHLBI
NIH HHS/United States Comparative Study Research Support, U.S. Gov't,
P.H.S. United states Proceedings of the National Academy of Sciences
of the United States of America Proc Natl Acad Sci U S A. 1991 Jul
15;88(14):6201-4.
%0 Journal Article
%1 Roth1991
%A Roth, N. S.
%A Campbell, P. T.
%A Caron, M. G.
%A Lefkowitz, R. J.
%A Lohse, M. J.
%D 1991
%J Proc Natl Acad Sci U S A
%K *Cyclic A/pharmacology AMP-Dependent Carcinoma, Cell Concanavalin GTP-Binding Humans Isoproterenol/*pharmacology Kinases Kinases/*metabolism Kinetics Line Membrane Permeability Protein Proteins/metabolism Receptor Squamous beta-Adrenergic beta/drug effects/*metabolism Adrenergic
%N 14
%P 6201-4
%T Comparative rates of desensitization of beta-adrenergic receptors
by the beta-adrenergic receptor kinase and the cyclic AMP-dependent
protein kinase
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1648731
%V 88
%X Three separate processes may contribute to rapid beta-adrenergic receptor
desensitization: functional uncoupling from the stimulatory guanine
nucleotide-binding protein Gs, mediated by phosphorylation of the
receptors by two distinct kinases, the specific beta-adrenergic receptor
kinase (beta ARK) and the cyclic AMP-dependent protein kinase A (PKA),
as well as a spatial uncoupling via sequestration of the receptors
away from the cell surface. To evaluate the relative importance and
potential role of the various processes in different physiological
situations, a kinetic analysis of these three mechanisms was performed
in permeabilized A431 epidermoid carcinoma cells. To allow a separate
analysis of each mechanism, inhibitors of the various desensitization
mechanisms were used: heparin to inhibit beta ARK, the PKA inhibitor
peptide PKI to inhibit PKA, and concanavalin A treatment to prevent
sequestration. Isoproterenol-induced phosphorylation of beta 2 receptors
in these cells by beta ARK occurred with a t1/2 of less than 20 sec,
whereas phosphorylation by PKA had a t1/2 of about 2 min. Similarly,
beta ARK-mediated desensitization of the receptors proceeded with
a t1/2 of less than 15 sec, and PKA-mediated desensitization with
a t1/2 of about 3.5 min. Maximal desensitization mediated by the
two kinases corresponded to a reduction of the signal-transduction
capacity of the receptor/adenylyl cyclase system by about 60% in
the case of beta ARK and by about 40% in the case of PKA. Receptor
sequestration was much slower (t1/2 of about 10 min) and involved
no more than 30% of the cell surface receptors. It is concluded that
beta ARK-mediated phosphorylation is the most rapid and quantitatively
most important factor contributing to the rapid desensitization.
This rapidity of the beta ARK-mediated mechanism makes it particularly
well suited to regulate beta-adrenergic receptor function in rapidly
changing environments such as the synaptic cleft.
@article{Roth1991,
abstract = {Three separate processes may contribute to rapid beta-adrenergic receptor
desensitization: functional uncoupling from the stimulatory guanine
nucleotide-binding protein Gs, mediated by phosphorylation of the
receptors by two distinct kinases, the specific beta-adrenergic receptor
kinase (beta ARK) and the cyclic AMP-dependent protein kinase A (PKA),
as well as a spatial uncoupling via sequestration of the receptors
away from the cell surface. To evaluate the relative importance and
potential role of the various processes in different physiological
situations, a kinetic analysis of these three mechanisms was performed
in permeabilized A431 epidermoid carcinoma cells. To allow a separate
analysis of each mechanism, inhibitors of the various desensitization
mechanisms were used: heparin to inhibit beta ARK, the PKA inhibitor
peptide PKI to inhibit PKA, and concanavalin A treatment to prevent
sequestration. Isoproterenol-induced phosphorylation of beta 2 receptors
in these cells by beta ARK occurred with a t1/2 of less than 20 sec,
whereas phosphorylation by PKA had a t1/2 of about 2 min. Similarly,
beta ARK-mediated desensitization of the receptors proceeded with
a t1/2 of less than 15 sec, and PKA-mediated desensitization with
a t1/2 of about 3.5 min. Maximal desensitization mediated by the
two kinases corresponded to a reduction of the signal-transduction
capacity of the receptor/adenylyl cyclase system by about 60% in
the case of beta ARK and by about 40% in the case of PKA. Receptor
sequestration was much slower (t1/2 of about 10 min) and involved
no more than 30% of the cell surface receptors. It is concluded that
beta ARK-mediated phosphorylation is the most rapid and quantitatively
most important factor contributing to the rapid desensitization.
This rapidity of the beta ARK-mediated mechanism makes it particularly
well suited to regulate beta-adrenergic receptor function in rapidly
changing environments such as the synaptic cleft.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Roth, N. S. and Campbell, P. T. and Caron, M. G. and Lefkowitz, R. J. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/2e3dd4af97f8fcdb96042b2c5339e70f0/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {0dc9b238e41c309c8f696eccfc579658},
intrahash = {e3dd4af97f8fcdb96042b2c5339e70f0},
issn = {0027-8424 (Print) 0027-8424 (Linking)},
journal = {Proc Natl Acad Sci U S A},
keywords = {*Cyclic A/pharmacology AMP-Dependent Carcinoma, Cell Concanavalin GTP-Binding Humans Isoproterenol/*pharmacology Kinases Kinases/*metabolism Kinetics Line Membrane Permeability Protein Proteins/metabolism Receptor Squamous beta-Adrenergic beta/drug effects/*metabolism Adrenergic},
month = {Jul 15},
note = {Roth, N S Campbell, P T Caron, M G Lefkowitz, R J Lohse, M J HL16037/HL/NHLBI
NIH HHS/United States Comparative Study Research Support, U.S. Gov't,
P.H.S. United states Proceedings of the National Academy of Sciences
of the United States of America Proc Natl Acad Sci U S A. 1991 Jul
15;88(14):6201-4.},
number = 14,
pages = {6201-4},
shorttitle = {Comparative rates of desensitization of beta-adrenergic receptors
by the beta-adrenergic receptor kinase and the cyclic AMP-dependent
protein kinase},
timestamp = {2010-12-14T18:22:42.000+0100},
title = {Comparative rates of desensitization of beta-adrenergic receptors
by the beta-adrenergic receptor kinase and the cyclic AMP-dependent
protein kinase},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1648731},
volume = 88,
year = 1991
}