Prostaglandin F2alpha-activated protein kinase Calpha phosphorylates myristoylated alanine-rich C kinase substrate protein in bovine luteal cells.
Prostaglandin F2alpha (PGF2alpha)-induced secretion of oxytocin by the bovine corpus luteum involves the phosphorylation of a unique protein kinase C (PKC) substrate, myristoylated alanine-rich C kinase substrate (MARCKS) protein. This study was conducted to determine the specific PKC isoform engaged in phosphorylation of MARCKS protein in bovine luteal cells. In experiment 1, dispersed luteal cells recovered from the corpus luteum on d 8 of the estrous cycle were preincubated with [32P] orthophosphate and then exposed to PGF2alpha alone or in combination with PKC inhibitors. Autoradiography and densitometry of Western blots revealed that MARCKS protein was phosphorylated by a conventional PKC (cPKC) isoform. Experiment 2 was conducted to identify the specific cPKC isoform that phosphorylates MARCKS protein in luteal cells. Corpora lutea were removed from control and PGF2alpha-treated heifers on d 8 of the cycle, and PKC isoforms associated with membrane and cytosolic fractions were determined. Treatment with PGF2alpha increased membrane concentrations of PKCalpha within 5 min after treatment (p < 0.005). Collectively, these data suggest that phosphorylation of MARCKS protein coinciding with oxytocin secretion is mediated by PKCalpha.
Salli
U
U
Department of Biochemistry/Biophysics, Oregon State University, Corvallis 97331, USA.
Stormshak
F
F
D000818:Animals; D002417:Cattle; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D015237:Dinoprost / Q000494:pharmacology*; D004789:Enzyme Activation / Q000502:physiology; D005260:Female; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes / Q000378:metabolism*; D008565:Membrane Proteins; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D011506:Proteins / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes; D008565:Membrane Proteins; D011506:Proteins; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D015237:Dinoprost; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha