Protein kinase C isoforms beta 1 and beta 2 inhibit the tyrosine kinase activity of the insulin receptor.
Downregulation of insulin receptor tyrosine kinase (IRK) activity yields to impaired insulin signalling and contributes to the pathogenesis of cellular insulin resistance. Activation of protein kinase C (PKC) by different agents is associated with an inhibition of IRK activity in various cell types. There is evidence that this effect on IRK activity might be mediated through phosphorylation of specific serine residues of the insulin receptor beta-subunit. Neither the domains of the IRK where inhibiting serine phosphorylation occurs nor the PKC isoform responsible for IRK inhibition have been identified. PKC consists of a family of at least 12 isoforms. The aim of the present study was to determine which PKC isoform might be capable of IRK inhibition. The human insulin receptor and the PKC isoforms alpha, beta 1, beta 2, gamma, delta, epsilon, eta, theta and zeta were overexpressed in human embryo kidney fibroblasts (HEK 293 cells) in order to answer this question. PKCs were activated by preincubation with the phorbolester (TPA) (10(-7) mol/l) following insulin stimulation of the cells. When the IRK was coexpressed with the PKC isoforms beta 1 and beta 2, a 50 +/- 15.7 and 45 +/- 10.1% inhibition of tyrosine autophosphorylation of IRK was observed while coexpression with the other isoforms did not significantly modify IRK autophosphorylation. The data suggest that the PKC isoforms beta 1 and beta 2 might be candidates for insulin receptor inhibition.
Bossenmaier
B
B
Hagedorn Research Institute, Gentofte, Denmark.
Mosthaf
L
L
Mischak
H
H
Ullrich
A
A
Häring
H U
HU
D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D007668:Kidney; D010766:Phosphorylation; D011493:Protein Kinase C / Q000096:biosynthesis / Q000378:metabolism*; D064546:Protein Kinase C beta; D011972:Receptor, Insulin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D011994:Recombinant Proteins; D011972:Receptor, Insulin; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D013755:Tetradecanoylphorbol Acetate