Mechanism of impaired metabolic signaling by a truncated human insulin receptor. Decreased activation of protein phosphatase 1 by insulin.
Previous studies have shown that a human insulin receptor lacking the COOH-terminal 43-amino acid domain (HIR delta CT) displays a compromised ability to stimulate glucose transport and glycogen synthase, whereas mitogenic signaling and stimulation of the insulin receptor tyrosine kinase activity remain intact (Maegawa, H., McClain, D. A., Freidenberg, G., Olefsky, J. M., Napier, M., Lipari, T., Dull, T. J., Lee, J., and Ullrich, A. (1988) J. Biol. Chem. 263, 8912-8917). In this study, we examined the effect of insulin on protein phosphatase 1 (PP-1) activity and phosphorylation in cells expressing wild-type human insulin receptor (HIRc) and HIR delta CT cells using phosphorylase alpha as substrate in the presence of 3 nM okadaic acid. Basal PP-1 activity was significantly lower in HIR delta CT than in HIRc cells (p < 0.05). Insulin stimulated PP-1 activity in HIRc cells (25-30% increase over basal activity) in a time- and dose-dependent manner. Insulin failed to stimulate PP-1 activity in HIR delta CT cells. Western blotting with the catalytic subunit antibody and the regulatory subunit antibody revealed similar amounts of the 37-kDa band (catalytic subunit) and the 160-kDa band (presumed regulatory subunit) in HIRc and HIR delta CT cells. We conclude that the COOH-terminal domain of the insulin receptor is an important element in mediating the effect of insulin on PP-1 and suggest that activation of PP-1 may be linked to signaling insulin's metabolic actions.
Department of Medicine, Veterans Affairs Medical Center, Denver, Colorado 80220.
N
N
Begum
J M
JM
Olefsky
B
B
Draznin
D000818:Animals; D002460:Cell Line; D003847:Deoxyglucose / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism*; D054645:Protein Phosphatase 1; D051381:Rats; D011972:Receptor, Insulin / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
D007328:Insulin; D011994:Recombinant Proteins; D003847:Deoxyglucose; D011972:Receptor, Insulin; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1