Regulation of angiotensin II-induced G protein signaling by phosducin-like protein.
Phosducin-like protein (PhLP) is a broadly expressed member of the phosducin (Pd) family of G protein betagamma subunit (Gbetagamma)-binding proteins. Though PhLP has been shown to bind Gbetagamma in vitro, little is known about its physiological function. In the present study, the effect of PhLP on angiotensin II (Ang II) signaling was measured in Chinese hamster ovary cells expressing the type 1 Ang II receptor and various amounts of PhLP. Up to 3.6-fold overexpression of PhLP had no effect on Ang II-stimulated inositol trisphosphate (IP(3)) formation, whereas further increases caused an abrupt decrease in IP(3) production with half-maximal inhibition occurring at 6-fold PhLP overexpression. This threshold level for inhibition corresponds to the cellular concentration of cytosolic chaperonin complex, a recently described binding partner that preferentially binds PhLP over Gbetagamma. Results of pertussis toxin sensitivity, GTPgammaS binding, and immunoprecipitation experiments suggest that PhLP inhibits phospholipase Cbeta activation by dual mechanisms: (i) steric blockage of Gbetagamma activation of PLCbeta and (ii) interference with Gbetagamma-dependent cycling of G(q)alpha by the receptor. These results suggest that G protein signaling may be regulated through controlling the cellular concentration of free PhLP by inducing its expression or by regulating its binding to the chaperonin.
McLaughlin
Joseph N
JN
Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, USA.
Thulin
Craig D
CD
Bray
Steven M
SM
Martin
Mickey M
MM
Elton
Terry S
TS
Willardson
Barry M
BM
D000804:Angiotensin II / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000502:physiology*; D006224:Cricetinae; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D007527:Isoenzymes / Q000378:metabolism; D018832:Molecular Chaperones; D009419:Nerve Tissue Proteins / Q000502:physiology*; D054799:Phospholipase C beta; D011233:Precipitin Tests; D051381:Rats; D011945:Receptors, Angiotensin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D010738:Type C Phospholipases / Q000378:metabolism
D002352:Carrier Proteins; D007527:Isoenzymes; D018832:Molecular Chaperones; D009419:Nerve Tissue Proteins; C498320:Pdcl protein, rat; D011945:Receptors, Angiotensin; D000804:Angiotensin II; D015544:Inositol 1,4,5-Trisphosphate; D010738:Type C Phospholipases; D054799:Phospholipase C beta; D019204:GTP-Binding Proteins