Expression of the insulin receptor with a recombinant vaccinia virus: Biochemical evidence that the insulin receptor has intrinsic serine kinase activity

Thomas J. Tauer, Deanna J. Volle, Solon L. Rhode, Robert E Lewis

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Abstract

We have previously reported the tight association of a serine kinase activity with the human insulin receptor (Lewis, R. E., Wu, G. P., MacDonald, R G., and Czech, M. P. (1990) J. Biol. Chem. 265, 947-954). We tested the possibility that the associated serine kinase activity was intrinsic to the receptor catalytic domain. The ratio of phosphoserine to phosphotyrosine on insulin receptors phosphorylated in vitro was used as an index of the associated serine kinase activity. Phosphorylation and phosphoamino acid analysis of insulin proreceptors revealed associated serine kinase activity early in receptor synthesis. Insulin receptors were expressed in HeLa cells using a recombinant vaccinia virus. The ratio of phosphoserine to phosphotyrosine on insulin receptors expressed by the recombinant vaccinia virus was determined relative to endogenous insulin receptors in cells treated with α-amanitin to block host cell mRNA synthesis. α-Amanitin treatment had no effect on the ratio of phosphoserine to phosphotyrosine on insulin receptors expressed from the recombinant virus even though they were present in a 4000-fold excess above endogenous receptors. We conclude that the serine kinase activity associated with the insulin receptor is intrinsic to the receptor catalytic domain. Receptor-catalyzed autophosphorylation of serine may play an important role in modulating insulin receptor signaling.

Original languageEnglish (US)
Pages (from-to)331-336
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number1
DOIs
StatePublished - Jan 5 1996

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Vaccinia virus
Protein-Serine-Threonine Kinases
Insulin Receptor
Viruses
Phosphoserine
Phosphotyrosine
Amanitins
Catalytic Domain
Phosphoamino Acids
Phosphorylation
HeLa Cells
Serine
Association reactions
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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Expression of the insulin receptor with a recombinant vaccinia virus : Biochemical evidence that the insulin receptor has intrinsic serine kinase activity. / Tauer, Thomas J.; Volle, Deanna J.; Rhode, Solon L.; Lewis, Robert E.

In: Journal of Biological Chemistry, Vol. 271, No. 1, 05.01.1996, p. 331-336.

Research output: Contribution to journalArticle

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