Regulation of the insulin receptor by protein kinase C isoenzymes: preferential interaction with beta isoenzymes and interaction with the catalytic domain of betaII.
We analysed the effects of high glucose in rat1 cells overexpressing insulin receptor. High (25 mM) glucose inhibited insulin-stimulated tyrosine kinase activity completely at insulin concentrations of 1 and 5 ng/ml. Decapeptides modelled on insulin receptor sequences surrounding serines 1035 and 1270 were found to inhibit protein kinase C activity in vitro and after microinjection into cells blocked the inhibition of mitogenesis induced by glucose. Purification of receptor from 3T3L1 adipocytes revealed that only the isoenzymes beta1, betaII and delta were detected. The site of the interaction was mapped to the catalytic domain of betaII. These results demonstrate that the inhibition of insulin receptor tyrosine kinase activity can be ameliorated using insulin receptor peptide sequences and there is constitutive and differential interaction of individual PKC isoenzymes with the insulin receptor, and in the case of betaII, this interaction maps to the catalytic domain rather than the regulatory domain.
Pillay
Tahir S
TS
Division of Endocrinology and Metabolism, Department of Medicine, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0673, USA. tpillay@nottingham.ac.uk
Xiao
Sen
S
Keranen
Lisa
L
Olefsky
Jerrold M
JM
D000818:Animals; D020134:Catalytic Domain / Q000502:physiology; D005947:Glucose / Q000378:metabolism*; D006943:Hyperglycemia / Q000378:metabolism* / Q000503:physiopathology; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D010446:Peptide Fragments / Q000494:pharmacology; D010766:Phosphorylation; D011485:Protein Binding / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011972:Receptor, Insulin / Q000378:metabolism*; D012694:Serine / Q000378:metabolism
D007328:Insulin; D007527:Isoenzymes; D010446:Peptide Fragments; D012694:Serine; C497259:Prkcd protein, mouse; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta; D005947:Glucose