Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor

Edward P. Feener, Jonathan M. Backer, George L. King, Peter A. Wilden, Xiao Jian Sun, C. Ronald Kahn, Morris F. White

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Insulin-stimulated autophosphorylation of the cytoplasmic juxtamembrane region of the human insulin receptor was examined by Tricine/SDS-PAGE. Various mutant receptor molecules were used to identify two tryptic phosphopeptides associated with the juxtamembrane region which accounts for 15% of the autophosphorylation of partially purified insulin receptor. These phosphopeptides were immunoprecipitated with an antipeptide antibody against the juxtamembrane sequence and were phosphorylated exclusively on tyrosine. Substitution of both Tyr960 and Tyr953 with alanine eliminated insulin-stimulated phosphorylation of the juxtamembrane region without affecting tyrosine autophosphorylation in the C terminus or regulatory regions. Monosubstitution of Tyr960 with phenylalanine or alanine reduced phosphorylation in the juxtamembrane region by more than 50%, and manual Edman degradation indicated that Tyr960 was phosphorylated in wild-type receptor. In vivo, phosphorylation of the juxtamembrane region accounts for one-third of the insulin receptor phosphorylation and contains both phosphoserine and phosphotyrosine. Deletion of Tyr960 and 11 adjacent amino acids eliminated insulin-stimulated phosphorylation of the juxtamembrane region. Substitution of Tyr960 reduced this phosphorylation by more than 50%. The insulin receptor also undergoes serine phosphorylation outside of the juxtamembrane region which depends on the presence of Tyr1151. Together with our previous studies, this report suggests that phosphorylation of Tyr960 may play an important role in signal transduction by the insulin receptor.

Original languageEnglish (US)
Pages (from-to)11256-11264
Number of pages9
JournalJournal of Biological Chemistry
Volume268
Issue number15
StatePublished - May 25 1993
Externally publishedYes

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Phosphorylation
Insulin Receptor
Serine
Tyrosine
Insulin
Phosphopeptides
Alanine
Substitution reactions
Phosphoserine
Signal transduction
Phosphotyrosine
Nucleic Acid Regulatory Sequences
Phenylalanine
Polyacrylamide Gel Electrophoresis
Signal Transduction
Amino Acids
Degradation
Molecules
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Feener, E. P., Backer, J. M., King, G. L., Wilden, P. A., Sun, X. J., Kahn, C. R., & White, M. F. (1993). Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor. Journal of Biological Chemistry, 268(15), 11256-11264.

Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor. / Feener, Edward P.; Backer, Jonathan M.; King, George L.; Wilden, Peter A.; Sun, Xiao Jian; Kahn, C. Ronald; White, Morris F.

In: Journal of Biological Chemistry, Vol. 268, No. 15, 25.05.1993, p. 11256-11264.

Research output: Contribution to journalArticle

Feener, EP, Backer, JM, King, GL, Wilden, PA, Sun, XJ, Kahn, CR & White, MF 1993, 'Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor', Journal of Biological Chemistry, vol. 268, no. 15, pp. 11256-11264.
Feener, Edward P. ; Backer, Jonathan M. ; King, George L. ; Wilden, Peter A. ; Sun, Xiao Jian ; Kahn, C. Ronald ; White, Morris F. / Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 15. pp. 11256-11264.
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