Cytoplasmic Juxtamembrane Region of the Insulin Receptor: A Critical Role in ATP Binding, Endogenous Substrate Phosphorylation, and Insulin-Stimulated Bioeffects in CHO Cells

Jonathan M. Backer, Gregory G. Schroeder, Deborah A. Cahill, Kenneth Siddle, Morris F. White, Axel Ullrich

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

We have expressed in CHO cells a mutant receptor (IRΔ960) from which 12 amino acids in the juxtamembrane region (A954-D965), including Tyr960, have been deleted. The mutant receptor bound insulin normally but exhibited an increased Km for ATP during autophosphorylation. Upon prolonged incubation in vitro, or at high ATP concentrations such as those observed in vivo, autophosphorylation of IRΔ960 was similar to wild type, and the in vitro phosphotransferase activity of the autophosphorylated IRΔ960 was normal. These results suggest that the deletion did not cause a nonspecific structural disruption of the catalytic domain of IRΔ960. In vivo autophosphorylation of the IRΔ960 receptor was reduced by 30% after 2 min of insulin stimulation and was similar to the wild-type receptor after 30 min of insulin stimulation. However, the mutant receptor was defective in insulin-stimulated tyrosyl phosphorylation of the endogenous substrate pp185. In additon, IRΔ960 was deficient in mediating insulin stimulation of glycogen and DNA synthesis. Thus, autophosphorylation of the insulin receptor is necessary but not sufficient for signal transmission. These data extend the hypothesis that the cytoplasmic juxtamembrane region of the insulin receptor is important for its interactions with ATP, intracellular substrates, and other proteins and is broadly necessary for biological signal transmission.

Original languageEnglish (US)
Pages (from-to)6366-6372
Number of pages7
JournalBiochemistry
Volume30
Issue number26
DOIs
StatePublished - Jul 1 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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