%0 Journal Article %1 Schroder1996 %A Schroder, S. %A Lohse, M. J. %D 1996 %J Proc Natl Acad Sci U S A %K & Alignment Animals Base Bordetella/antagonists Carrier DNA Data Enzyme Eye Factors, GTP GTP-Binding Inhibitors Molecular Nerve Pertussis Phosphohydrolases/antagonists Phosphoproteins/*chemistry Phosphorylation Primers/chemistry Protein Proteins Proteins/*antagonists Proteins/*chemistry Proteins/isolation Rats Recombinant Regulators Rhodopsin/metabolism Sequence Signal Tissue Toxin Transduction Virulence inhibitors purification/*metabolism %N 5 %P 2100-4 %T Inhibition of G-protein betagamma-subunit functions by phosducin-like protein %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8700891 %V 93 %X Phosducin is a cytosolic protein predominantly expressed in the retina and the pineal gland that can interact with the betagamma subunits of guanine nucleotide binding proteins (G proteins) and thereby may regulate transmembrane signaling. A cDNA encoding a phosducin-like protein (PhLP) has recently been isolated from rat brain Miles, M. F., Barhite, S., Sganga, M. & Elliott, M. (1993) Proc. Natl. Acad. Sci. USA 90, 10831-10835. Here we report the expression of PhLP in Escherichia coli and its purification. Recombinant purified PUP inhibited multiple effects of G-protein betagamma subunits. First, it inhibited the betagamma-subunit-dependent ADP-ribosylation of purified alpha(o) by pertussis toxin. Second, it inhibited the GTPase activity of purified G(o). The IC50 value of PhLP in the latter assay was 89 nM, whereas phosducin caused half-maximal inhibition at 17 nM. And finally, PhLP antagonized the enhancement of rhodopsin phosphorylation by purified betagamma subunits. The N terminus of PhLP shows no similarity to the much longer N terminus of phosducin, the region shown to be critical for phosducin-betagamma-subunit interactions. Therefore, PhLP appears to bind to G-protein betagamma subunits by an as yet unknown mode of interaction and may represent an endogenous regulator of G-protein function.

@article{Schroder1996, abstract = {Phosducin is a cytosolic protein predominantly expressed in the retina and the pineal gland that can interact with the betagamma subunits of guanine nucleotide binding proteins (G proteins) and thereby may regulate transmembrane signaling. A cDNA encoding a phosducin-like protein (PhLP) has recently been isolated from rat brain [Miles, M. F., Barhite, S., Sganga, M. & Elliott, M. (1993) Proc. Natl. Acad. Sci. USA 90, 10831-10835. Here we report the expression of PhLP in Escherichia coli and its purification. Recombinant purified PUP inhibited multiple effects of G-protein betagamma subunits. First, it inhibited the betagamma-subunit-dependent ADP-ribosylation of purified alpha(o) by pertussis toxin. Second, it inhibited the GTPase activity of purified G(o). The IC50 value of PhLP in the latter assay was 89 nM, whereas phosducin caused half-maximal inhibition at 17 nM. And finally, PhLP antagonized the enhancement of rhodopsin phosphorylation by purified betagamma subunits. The N terminus of PhLP shows no similarity to the much longer N terminus of phosducin, the region shown to be critical for phosducin-betagamma-subunit interactions. Therefore, PhLP appears to bind to G-protein betagamma subunits by an as yet unknown mode of interaction and may represent an endogenous regulator of G-protein function.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Schroder, S. and Lohse, M. J.}, biburl = {https://www.bibsonomy.org/bibtex/21371082ad928e016d4fce5e0a9507a44/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {2181d280c1c5d6a47e7b6294237e7bb3}, intrahash = {1371082ad928e016d4fce5e0a9507a44}, issn = {0027-8424 (Print) 0027-8424 (Linking)}, journal = {Proc Natl Acad Sci U S A}, keywords = {& Alignment Animals Base Bordetella/antagonists Carrier DNA Data Enzyme Eye Factors, GTP GTP-Binding Inhibitors Molecular Nerve Pertussis Phosphohydrolases/antagonists Phosphoproteins/*chemistry Phosphorylation Primers/chemistry Protein Proteins Proteins/*antagonists Proteins/*chemistry Proteins/isolation Rats Recombinant Regulators Rhodopsin/metabolism Sequence Signal Tissue Toxin Transduction Virulence inhibitors purification/*metabolism}, month = {Mar 5}, note = {Schroder, S Lohse, M J Research Support, Non-U.S. Gov't United states Proceedings of the National Academy of Sciences of the United States of America Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):2100-4.}, number = 5, pages = {2100-4}, shorttitle = {Inhibition of G-protein betagamma-subunit functions by phosducin-like protein}, timestamp = {2010-12-14T18:12:31.000+0100}, title = {Inhibition of G-protein betagamma-subunit functions by phosducin-like protein}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8700891}, volume = 93, year = 1996 }