Transmembrane signaling by the human insulin receptor kinase: Relationship between intramolecular β subunit trans- and cis-autophosphorylation and substrate kinase activation

Anne L. Frattali, Judith L. Treadway, Jeffrey E. Pessin

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Abstract

To examine the role of intramolecular β subunit trans- and cis-autophosphorylation in signal transduction, the vaccinia virus/bacteriophage T7 expression system was used to generate insulin holoreceptors composed of a kinase-defective half-receptor precursor (αβA/K or αβA/K.ΔCT) and a kinase-active half-receptor precursor (αβΔCT or αβWT). In the αβWT-αβΔCT hybrid insulin receptor, insulin stimulated a 20-fold increase in intramolecular β subunit trans-phosphorylation, whereas cis-phosphorylation increased only 3-fold over the basal state. Similarly, in the αβWT-αβA/K.ΔCT hybrid insulin receptor, insulin stimulated trans-phosphorylation approximately 30-fold and cis-phosphorylation only 3-fold over the basal state. Although cis-phosphorylation of the kinase-functional αβ half-receptor was observed within these hybrid receptor species, this was not sufficient to stimulate exogenous substrate kinase activity. These data demonstrate that insulin primarily activates an intramolecular β subunit trans-phosphorylation reaction within the insulin holoreceptor and suggest that this reaction is necessary for activation of the holoreceptor. Furthermore, our results suggest a molecular basis for the dominant-negative phenotype observed in insulin-resistant patients possessing one kinase-defective insulin receptor allele.

Original languageEnglish (US)
Pages (from-to)19521-19528
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number27
StatePublished - Sep 25 1992
Externally publishedYes

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Phosphorylation
Phosphotransferases
Chemical activation
Insulin
Insulin Receptor
Substrates
Bacteriophage T7
Signal transduction
Bacteriophages
Vaccinia virus
human INSR protein
Signal Transduction
Alleles
Phenotype

ASJC Scopus subject areas

  • Biochemistry

Cite this

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abstract = "To examine the role of intramolecular β subunit trans- and cis-autophosphorylation in signal transduction, the vaccinia virus/bacteriophage T7 expression system was used to generate insulin holoreceptors composed of a kinase-defective half-receptor precursor (αβA/K or αβA/K.ΔCT) and a kinase-active half-receptor precursor (αβΔCT or αβWT). In the αβWT-αβΔCT hybrid insulin receptor, insulin stimulated a 20-fold increase in intramolecular β subunit trans-phosphorylation, whereas cis-phosphorylation increased only 3-fold over the basal state. Similarly, in the αβWT-αβA/K.ΔCT hybrid insulin receptor, insulin stimulated trans-phosphorylation approximately 30-fold and cis-phosphorylation only 3-fold over the basal state. Although cis-phosphorylation of the kinase-functional αβ half-receptor was observed within these hybrid receptor species, this was not sufficient to stimulate exogenous substrate kinase activity. These data demonstrate that insulin primarily activates an intramolecular β subunit trans-phosphorylation reaction within the insulin holoreceptor and suggest that this reaction is necessary for activation of the holoreceptor. Furthermore, our results suggest a molecular basis for the dominant-negative phenotype observed in insulin-resistant patients possessing one kinase-defective insulin receptor allele.",
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AU - Pessin, Jeffrey E.

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