Arachidonic acid-induced insulin secretion from rat islets of Langerhans is not mediated by protein phosphorylation.
Arachidonic acid (AA) stimulated protein phosphorylation in electrically permeabilised islets, most notably of an islet protein of approximate molecular weight 18 kDa. This protein did not appear to be a substrate for cAMP-dependent protein kinase. The AA-induced protein phosphorylation was mediated by unmetabolised AA since the lipoxygenase inhibitor, nordihydroguaretic acid (NDGA), or the cyclooxygenase inhibitor, indomethacin, did not significantly reduce AA-induced phosphorylation. Although saturated fatty acids did not stimulate phosphorylation of islet proteins, a number of cis-unsaturated fatty acids, other than AA, induced 32P incorporation into an 18 kDa protein. However, some fatty acids which stimulated protein phosphorylation had no effect on insulin secretion in experiments where AA clearly stimulated insulin secretion. AA stimulated protein kinase C (PKC) activity extracted from islets but several fatty acids which induced protein phosphorylation had no significant effect on PKC activity in vitro. 50 nM staurosporine had no effect on AA-induced protein phosphorylation but this concentration of staurosporine markedly inhibited PKC activity. 200 nM staurosporine caused complete inhibition of the AA-induced phosphorylation without having any effect on AA-induced insulin secretion. These results suggest that AA and some other fatty acids can promote 32P incorporation into islet proteins, independently of PKC activation, and that AA-induced phosphorylation is not required for insulin secretory responses to AA.
Biomedical Sciences Division, King's College London, UK.
H
H
Basudev
P M
PM
Jones
S J
SJ
Persaud
S L
SL
Howell
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology*; D000242:Cyclic AMP / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D009637:Masoprocol / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D019311:Staurosporine
D000470:Alkaloids; D005231:Fatty Acids, Unsaturated; D007328:Insulin; D010750:Phosphoproteins; D016718:Arachidonic Acid; D009637:Masoprocol; D000242:Cyclic AMP; D011494:Protein Kinases; D011493:Protein Kinase C; D019311:Staurosporine; D007213:Indomethacin