Insulin and ribosomal protein S6 kinase in rat pancreatic acini.
Treatment of pancreatic acini from diabetic rats with insulin resulted in a dose-dependent increase in the phosphorylation of ribosomal protein S6 when analyzed by two-dimensional gel electrophoresis. To study the presence of the protein kinase mediating this phosphorylation, soluble extracts of intact acini that had been previously treated with insulin were prepared and assayed for protein kinase activity with rat pancreatic ribosomes as a substrate. Activation of S6 kinase activity, observed in a time-dependent manner, was maximal after 20-30 min and, in a dose-dependent manner, was half-maximal at 1 nM and maximal at 10 nM insulin concentration. Based on cofactor requirements, substrate specificity, and a slow activation of the enzyme, the S6 kinase was distinct from cAMP-dependent, Ca2+-calmodulin-dependent, and Ca2+-phospholipid-dependent protein kinases and protease-activated kinase II. The S6 kinase activated by insulin was highly specific for the ribosomal protein S6 when compared with various substrates, including casein, glycogen synthase, phosphorylase b, phosvitin, histone HIII-S, and histone HVIII-S. Protein S6 phosphorylation in intact acini and activation of the S6 kinase by insulin showed similar dose-response curves, consistent with the S6 kinase being responsible for the protein S6 phosphorylation in intact acini. The comparison of the dose-response curves for S6 phosphorylation and protein synthesis in acini suggests that there is a close correlation between these two insulin actions.
Sung
C K
CK
Department of Physiology, University of California, San Francisco.
Williams
J A
JA
D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D007328:Insulin / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000187:drug effects / Q000201:enzymology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D038601:Ribosomal Protein S6; D019893:Ribosomal Protein S6 Kinases; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects / Q000201:enzymology
D007328:Insulin; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D011494:Protein Kinases; D019893:Ribosomal Protein S6 Kinases