Characterization of insulin-stimulated seryl/threonyl protein kinases in rat skeletal muscle.
Post-insulin receptor signal transduction is mediated by a cascade of seryl/threonyl protein kinases which includes a family of mitogen-activated protein (MAP) kinases, ribosomal protein S6 kinases, and casein kinase-2. Previous studies have characterized these kinases primarily in cultured or isolated cells. We have demonstrated that intravenous injection of insulin into fasted rats significantly stimulated the activities of MAP kinases and S6 kinases in skeletal muscle, independently of the blood glucose levels in these animals. Anion exchange chromatography on Mono Q afforded the resolution of at least five peaks of insulin-stimulated myelin basic protein kinase activity. By immunological criteria, these myelin basic protein kinases included the p42mapk and p44erk1 as well as other potentially novel 44-kDa MAP kinases. Insulin-activated ribosomal S6 kinases were resolved into two major peaks by Mono Q chromatography, the latter of which contained a 100-kDa isoform of p90rsk as revealed by immunoblotting with an anti-rsk-peptide antibody. A 32-kDa S6 kinase in the earlier peak may represent a novel protein kinase in this tissue. Skeletal muscle casein kinase-2 was not significantly stimulated following insulin injection into rats under our experimental conditions. These results indicate that the intact rat can serve as a useful model system to investigate the mechanisms of insulin signal transduction.
Hei
Y J
YJ
Division of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, Canada.
McNeill
J H
JH
Sanghera
J S
JS
Diamond
J
J
Bryer-Ash
M
M
Pelech
S L
SL
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002850:Chromatography, Gel; D004789:Enzyme Activation; D007328:Insulin / Q000494:pharmacology*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology*; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D013217:Starvation
D007328:Insulin; D007527:Isoenzymes; D017346:Protein Serine-Threonine Kinases