Identification of a C-terminal binding site for G-protein betagamma-subunits
in phosducin-like protein
S. Schroder, K. Bluml, C. Dees, and M. Lohse. FEBS Lett, 401 (2-3):
243-6(January 1997)Schroder, S Bluml, K Dees, C Lohse, M J Research Support, Non-U.S.
Gov't Netherlands FEBS letters FEBS Lett. 1997 Jan 20;401(2-3):243-6..
Abstract
Phosducin-like protein (PhLP) has recently been identified as a ubiquitous
inhibitor of G-protein betagamma-subunit (G betagamma)-mediated signaling,
with an affinity about 5-fold lower than that of phosducin. The G
betagamma binding site of phosducin has been suggested to be contained
in its N-terminus. A region corresponding to this N-terminus is lacking
in PhLP, suggesting that PhLP must utilize a different mode of G
betagamma binding. To map the G betagamma binding site in PhLP, a
series of deletion mutants were constructed, expressed in E. coli
as glutathione S-transferase (GST) fusion proteins, and the purified
fusion proteins were examined for their ability to attenuate G(o)
GTPase activity. Progressive N-terminal truncations of PhLP caused
only minor reductions in potency, whereas the complementary N-terminal
PhLP fragments turned out to be inactive. We further identified a
short C-terminal segment comprising residues 168 to 195 that inhibited
G0 GTPase activity similar in efficacy and potency to full-length
PhLP. This C-terminal fragment was also capable of antagonizing a
second G betagamma-mediated function, the enhancement of rhodopsin
phosphorylation by the beta-adrenergic receptor kinase. Taken together,
these data indicate that PhLP interacts with G betagamma via a short
C-terminal binding site which is distinct from that identified previously
in phosducin.
%0 Journal Article
%1 Schroder1997
%A Schroder, S.
%A Bluml, K.
%A Dees, C.
%A Lohse, M. J.
%D 1997
%J FEBS Lett
%K & Animals Binding Carrier Deletion Escherichia Fusion GTP GTP-Binding Nerve Phosphohydrolases/antagonists Proteins/antagonists Proteins/genetics/*metabolism Proteins/genetics/metabolism Recombinant Sequence Sites Tissue coli inhibitors/*metabolism inhibitors/metabolism
%N 2-3
%P 243-6
%T Identification of a C-terminal binding site for G-protein betagamma-subunits
in phosducin-like protein
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9013896
%V 401
%X Phosducin-like protein (PhLP) has recently been identified as a ubiquitous
inhibitor of G-protein betagamma-subunit (G betagamma)-mediated signaling,
with an affinity about 5-fold lower than that of phosducin. The G
betagamma binding site of phosducin has been suggested to be contained
in its N-terminus. A region corresponding to this N-terminus is lacking
in PhLP, suggesting that PhLP must utilize a different mode of G
betagamma binding. To map the G betagamma binding site in PhLP, a
series of deletion mutants were constructed, expressed in E. coli
as glutathione S-transferase (GST) fusion proteins, and the purified
fusion proteins were examined for their ability to attenuate G(o)
GTPase activity. Progressive N-terminal truncations of PhLP caused
only minor reductions in potency, whereas the complementary N-terminal
PhLP fragments turned out to be inactive. We further identified a
short C-terminal segment comprising residues 168 to 195 that inhibited
G0 GTPase activity similar in efficacy and potency to full-length
PhLP. This C-terminal fragment was also capable of antagonizing a
second G betagamma-mediated function, the enhancement of rhodopsin
phosphorylation by the beta-adrenergic receptor kinase. Taken together,
these data indicate that PhLP interacts with G betagamma via a short
C-terminal binding site which is distinct from that identified previously
in phosducin.
@article{Schroder1997,
abstract = {Phosducin-like protein (PhLP) has recently been identified as a ubiquitous
inhibitor of G-protein betagamma-subunit (G betagamma)-mediated signaling,
with an affinity about 5-fold lower than that of phosducin. The G
betagamma binding site of phosducin has been suggested to be contained
in its N-terminus. A region corresponding to this N-terminus is lacking
in PhLP, suggesting that PhLP must utilize a different mode of G
betagamma binding. To map the G betagamma binding site in PhLP, a
series of deletion mutants were constructed, expressed in E. coli
as glutathione S-transferase (GST) fusion proteins, and the purified
fusion proteins were examined for their ability to attenuate G(o)
GTPase activity. Progressive N-terminal truncations of PhLP caused
only minor reductions in potency, whereas the complementary N-terminal
PhLP fragments turned out to be inactive. We further identified a
short C-terminal segment comprising residues 168 to 195 that inhibited
G0 GTPase activity similar in efficacy and potency to full-length
PhLP. This C-terminal fragment was also capable of antagonizing a
second G betagamma-mediated function, the enhancement of rhodopsin
phosphorylation by the beta-adrenergic receptor kinase. Taken together,
these data indicate that PhLP interacts with G betagamma via a short
C-terminal binding site which is distinct from that identified previously
in phosducin.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Schroder, S. and Bluml, K. and Dees, C. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/29569cdc6543fcf7358fcbf13225617da/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {9b3dbf82c46599d7c5668981dcba744a},
intrahash = {9569cdc6543fcf7358fcbf13225617da},
issn = {0014-5793 (Print) 0014-5793 (Linking)},
journal = {FEBS Lett},
keywords = {& Animals Binding Carrier Deletion Escherichia Fusion GTP GTP-Binding Nerve Phosphohydrolases/antagonists Proteins/antagonists Proteins/genetics/*metabolism Proteins/genetics/metabolism Recombinant Sequence Sites Tissue coli inhibitors/*metabolism inhibitors/metabolism},
month = {Jan 20},
note = {Schroder, S Bluml, K Dees, C Lohse, M J Research Support, Non-U.S.
Gov't Netherlands FEBS letters FEBS Lett. 1997 Jan 20;401(2-3):243-6.},
number = {2-3},
pages = {243-6},
shorttitle = {Identification of a C-terminal binding site for G-protein betagamma-subunits
in phosducin-like protein},
timestamp = {2010-12-14T18:22:03.000+0100},
title = {Identification of a C-terminal binding site for G-protein betagamma-subunits
in phosducin-like protein},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9013896},
volume = 401,
year = 1997
}