Differential regulation of prostaglandin F(2alpha) receptor isoforms by protein kinase C.
Prostaglandin F(2alpha) receptors (FP) are G protein-coupled receptors that bind prostaglandin F(2alpha) (PGF(2alpha)), resulting in the activation of an inositol phosphate (IP) second messenger pathway. Alternative mRNA splicing generates two FP receptor isoforms. These isoforms, designated FP(A) and FP(B), are otherwise identical except for their carboxyl termini. FP(B) is essentially a truncated version of FP(A) that lacks the 46 carboxyl-terminal amino acids, including four putative protein kinase C (PKC) phosphorylation sites. Until now, functional differences between these FP receptor isoforms have not been identified. We now report that pretreatment with the PKC inhibitor bisindolylmaleimide I enhanced PGF(2alpha)-stimulated IP accumulation in transfected cells stably expressing the FP(A) isoform but not in cells stably expressing the FP(B) isoform. Whole-cell phosphorylation experiments showed a strong agonist-dependent phosphorylation of the FP(A) isoform but little or no phosphorylation of the FP(B). Pretreatment of cells with bisindolylmaleimide I decreased PGF(2alpha)-stimulated phosphorylation of the FP(A) isoform consistent with a PKC-dependent phosphorylation. In vitro phosphorylation of an FP(A) carboxyl-terminal fusion protein by recombinant PKCalpha showed that the carboxyl terminus of the FP(A) is a substrate for PKC. These results suggest that PKC-dependent phosphorylation is responsible for differential regulation of second messenger signaling by FP prostanoid receptor isoforms.
Fujino
H
H
Department of Pharmacology, College of Pharmacy, University of Arizona, Tucson, Arizona, USA.
Srinivasan
D
D
Pierce
K L
KL
Regan
J W
JW
D002460:Cell Line; D015237:Dinoprost / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D010766:Phosphorylation; D020033:Protein Isoforms / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism / Q000502:physiology*; D051571:Protein Kinase C-alpha; D011982:Receptors, Prostaglandin / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D007295:Inositol Phosphates; D007527:Isoenzymes; D020033:Protein Isoforms; D011982:Receptors, Prostaglandin; D011993:Recombinant Fusion Proteins; C034671:prostaglandin F2alpha receptor; D015237:Dinoprost; D005982:Glutathione Transferase; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha