Redox priming of the insulin receptor beta-chain associated with altered tyrosine kinase activity and insulin responsiveness in the absence of tyrosine autophosphorylation.
Induction of tyrosine kinase activity of the insulin receptor (IR) beta-chain is believed to require its autophosphorylation at Tyr1162, Tyr1163, and Tyr1158. However, the mechanism of the initial phosphorylation is poorly understood. We show that treatment of IR-transfected Chinese hamster ovary cells with antioxidants inhibits insulin responsiveness. Conversely, partial inhibition of glutathione biosynthesis by buthionine sulfoximine (BSO) and glutathione reductase by 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU), i.e., procedures that intracellularly induce mildly oxidative conditions, caused a decrease in IR beta-chain sulfhydryl groups and enhanced synergistically the induction of IR tyrosine phosphorylation by insulin. The IR beta-chain from cells treated with BSO/BCNU in the absence of insulin was not detectably tyrosine phosphorylated, but nevertheless was functionally altered, as demonstrated in vitro by a moderate kinase activity at lowATP concentrations (5 nM) and a strong kinase activity at 25 microM ATP. This activity was found to be specific for tyrosine (not for serine or threonine), and tryptic peptide maps indicated that it is more selective than that induced by insulin. Moreover, the kinase activity from BSO/BCNU-treated cells showed a spontaneous decay that was not prevented by the phosphatase inhibitor vanadate. Together, these results suggest that optimal insulin responsiveness may require a process of 'redox priming' of the IR beta-chain that involves structural and functional changes in the absence of detectable tyrosine phosphorylation of the beta-chain.
Schmid
E
E
Deutsches Krebsforschungszentrum, Division of Immunochemistry, Heidelberg, Germany.
El Benna
J
J
Galter
D
D
Klein
G
G
Dröge
W
W
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology*; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D019328:Buthionine Sulfoximine / Q000494:pharmacology*; D016466:CHO Cells / Q000187:drug effects; D002330:Carmustine / Q000494:pharmacology*; D006224:Cricetinae; D004790:Enzyme Induction; D007328:Insulin / Q000502:physiology*; D010084:Oxidation-Reduction; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D011972:Receptor, Insulin / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism*
D000963:Antimetabolites; D018906:Antineoplastic Agents, Alkylating; D000975:Antioxidants; D007328:Insulin; D014443:Tyrosine; D019328:Buthionine Sulfoximine; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D010744:Phosphoric Monoester Hydrolases; D002330:Carmustine