Expression of phosducin in a phosducin-negative cell line reveals
functions of a Gbetagamma-binding protein
K. Schulz, S. Danner, P. Bauer, S. Schroder, и M. Lohse. J Biol Chem, 271 (37):
22546-51(сентября 1996)Schulz, K Danner, S Bauer, P Schroder, S Lohse, M J Research Support,
Non-U.S. Gov't United states The Journal of biological chemistry
J Biol Chem. 1996 Sep 13;271(37):22546-51..
Аннотация
Phosducin is a member of the large group of proteins that bind to
G-protein betagamma-subunits (Gbetagamma) and whose biological functions
are often unknown. Human A431 cells do not contain detectable amounts
of phosducin. We generated A431 cells expressing phosducin at a level
of approximately 1 pmol/mg of cytosolic protein, which is approximately
10% of the phosducin level in brain. cAMP accumulation in response
to beta2-adrenergic receptor agonists was enhanced at early times
in phosducin-expressing cells, but reached a lower plateau than in
control cells. Permeabilization of the cells with digitonin did not
change this pattern, but allowed the introduction of specific inhibitors:
antibodies to phosducin abolished all differences between the two
cell lines. Inhibitors of the beta-adrenergic receptor kinase abolished
the differences at early time points. An almost complete loss of
beta2-adrenergic receptor desensitization in the phosducin-expressing
cells was also observed when intact cells were desensitized and receptor
function was then determined in membrane preparations. Inhibition
of protein kinase A accentuated the effects of phosducin, suggesting
that also in vivo phosducin is regulated by this kinase. These data
indicate that phosducin affects G-protein-mediated signaling in at
least two ways: it dampens the overall responsiveness, and it impairs
the rapid desensitization mediated by the beta-adrenergic receptor
kinase.
Schulz, K Danner, S Bauer, P Schroder, S Lohse, M J Research Support,
Non-U.S. Gov't United states The Journal of biological chemistry
J Biol Chem. 1996 Sep 13;271(37):22546-51.
%0 Journal Article
%1 Schulz1996
%A Schulz, K.
%A Danner, S.
%A Bauer, P.
%A Schroder, S.
%A Lohse, M. J.
%D 1996
%J J Biol Chem
%K & 1-Methyl-3-isobutylxanthine/pharmacology AMP-Dependent AMP/metabolism Acid Adenylate Adrenergic Amino Blotting, Cell Cyclase/metabolism Cyclic Data Eye GTP-Binding Humans Inhibitors/pharmacology Isoproterenol/pharmacology Kinases/antagonists Line Molecular Phosphodiesterase Phosphoproteins/*biosynthesis Protein Proteins/*biosynthesis Regulators Sequence Signal Transduction Transfection Western beta-Agonists/pharmacology beta/metabolism inhibitors Receptor Proteins/metabolism
%N 37
%P 22546-51
%T Expression of phosducin in a phosducin-negative cell line reveals
functions of a Gbetagamma-binding protein
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8798422
%V 271
%X Phosducin is a member of the large group of proteins that bind to
G-protein betagamma-subunits (Gbetagamma) and whose biological functions
are often unknown. Human A431 cells do not contain detectable amounts
of phosducin. We generated A431 cells expressing phosducin at a level
of approximately 1 pmol/mg of cytosolic protein, which is approximately
10% of the phosducin level in brain. cAMP accumulation in response
to beta2-adrenergic receptor agonists was enhanced at early times
in phosducin-expressing cells, but reached a lower plateau than in
control cells. Permeabilization of the cells with digitonin did not
change this pattern, but allowed the introduction of specific inhibitors:
antibodies to phosducin abolished all differences between the two
cell lines. Inhibitors of the beta-adrenergic receptor kinase abolished
the differences at early time points. An almost complete loss of
beta2-adrenergic receptor desensitization in the phosducin-expressing
cells was also observed when intact cells were desensitized and receptor
function was then determined in membrane preparations. Inhibition
of protein kinase A accentuated the effects of phosducin, suggesting
that also in vivo phosducin is regulated by this kinase. These data
indicate that phosducin affects G-protein-mediated signaling in at
least two ways: it dampens the overall responsiveness, and it impairs
the rapid desensitization mediated by the beta-adrenergic receptor
kinase.
@article{Schulz1996,
abstract = {Phosducin is a member of the large group of proteins that bind to
G-protein betagamma-subunits (Gbetagamma) and whose biological functions
are often unknown. Human A431 cells do not contain detectable amounts
of phosducin. We generated A431 cells expressing phosducin at a level
of approximately 1 pmol/mg of cytosolic protein, which is approximately
10% of the phosducin level in brain. cAMP accumulation in response
to beta2-adrenergic receptor agonists was enhanced at early times
in phosducin-expressing cells, but reached a lower plateau than in
control cells. Permeabilization of the cells with digitonin did not
change this pattern, but allowed the introduction of specific inhibitors:
antibodies to phosducin abolished all differences between the two
cell lines. Inhibitors of the beta-adrenergic receptor kinase abolished
the differences at early time points. An almost complete loss of
beta2-adrenergic receptor desensitization in the phosducin-expressing
cells was also observed when intact cells were desensitized and receptor
function was then determined in membrane preparations. Inhibition
of protein kinase A accentuated the effects of phosducin, suggesting
that also in vivo phosducin is regulated by this kinase. These data
indicate that phosducin affects G-protein-mediated signaling in at
least two ways: it dampens the overall responsiveness, and it impairs
the rapid desensitization mediated by the beta-adrenergic receptor
kinase.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Schulz, K. and Danner, S. and Bauer, P. and Schroder, S. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/24042eee7755a39659daf4ce6c0e2482a/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {ac968bcbd8b5d103d80603c1909abe10},
intrahash = {4042eee7755a39659daf4ce6c0e2482a},
issn = {0021-9258 (Print) 0021-9258 (Linking)},
journal = {J Biol Chem},
keywords = {& 1-Methyl-3-isobutylxanthine/pharmacology AMP-Dependent AMP/metabolism Acid Adenylate Adrenergic Amino Blotting, Cell Cyclase/metabolism Cyclic Data Eye GTP-Binding Humans Inhibitors/pharmacology Isoproterenol/pharmacology Kinases/antagonists Line Molecular Phosphodiesterase Phosphoproteins/*biosynthesis Protein Proteins/*biosynthesis Regulators Sequence Signal Transduction Transfection Western beta-Agonists/pharmacology beta/metabolism inhibitors Receptor Proteins/metabolism},
month = {Sep 13},
note = {Schulz, K Danner, S Bauer, P Schroder, S Lohse, M J Research Support,
Non-U.S. Gov't United states The Journal of biological chemistry
J Biol Chem. 1996 Sep 13;271(37):22546-51.},
number = 37,
pages = {22546-51},
shorttitle = {Expression of phosducin in a phosducin-negative cell line reveals
functions of a Gbetagamma-binding protein},
timestamp = {2010-12-14T18:22:42.000+0100},
title = {Expression of phosducin in a phosducin-negative cell line reveals
functions of a Gbetagamma-binding protein},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8798422},
volume = 271,
year = 1996
}