Insulin stimulates tyrosine phosphorylation of multiple high molecular weight substrates in Fao hepatoma cells

Montserrat Miralpeix, Xiao Jian Sun, Jonathan M. Backer, Martin G. Myers, Eichii Araki, Morris F. White

Research output: Contribution to journalArticle

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Abstract

Insulin rapidly stimulates tyrosine phosphorylation of cellular proteins which migrate between 165 and 190 kDa during SDS-PAGE. These proteins, collectively called pp185, were originally found in anti-phosphotyrosine antibody (αPY) immunoprecipitates from insulin-stimulated Fao rat hepatoma cells. Recently, we purified and cloned IRS-1, one of the phosphoproteins that binds to αPY and migrates near 180 kDa following insulin stimulation of rat liver [Sun, X. J., et al. (1991) Nature 352, 73-77]. IRS-1 and pp185 undergo tyrosine phosphorylation immediately after insulin stimulation and show an insulin dose response similar to that of insulin receptor autophosphorylation. However, IRS-1 was consistently 10 kDa smaller than the apparent molecular mass of pp185. The pp185 contained some immunoblottable IRS-1; however, cell lysates depleted of IRS-1 with anti-IRS-1 antibody still contained the high molecular weight forms of pp185 (HMW-pp185). Furthermore, the tryptic phosphopeptide map of IRS-1 was distinct from that of HMW-pp185, suggesting that at least two substrates migrate in this region during SDS-PAGE. Moreover, the phosphatidylinositol 3′-kinase and its 85-kDa associated protein (p85) bound to IRS-1 in Fao cells, but weakly or not at all to HMW-pp185. Our results show that Fao cells contain at least two insulin receptor substrates, IRS-1 and HMW-pp185, which may play unique roles in insulin signal transmission.

Original languageEnglish (US)
Pages (from-to)9031-9039
Number of pages9
JournalBiochemistry
Volume31
Issue number37
StatePublished - 1992
Externally publishedYes

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Phosphorylation
Tyrosine
Hepatocellular Carcinoma
Molecular Weight
Molecular weight
Insulin
Substrates
Phosphotyrosine
Insulin Receptor
Rats
Polyacrylamide Gel Electrophoresis
Phosphatidylinositol 3-Kinase
Insulin Antibodies
Phosphopeptides
Proteins
Antibodies
Phosphoproteins
Insulin Receptor Substrate Proteins
Molecular mass
Liver

ASJC Scopus subject areas

  • Biochemistry

Cite this

Miralpeix, M., Sun, X. J., Backer, J. M., Myers, M. G., Araki, E., & White, M. F. (1992). Insulin stimulates tyrosine phosphorylation of multiple high molecular weight substrates in Fao hepatoma cells. Biochemistry, 31(37), 9031-9039.

Insulin stimulates tyrosine phosphorylation of multiple high molecular weight substrates in Fao hepatoma cells. / Miralpeix, Montserrat; Sun, Xiao Jian; Backer, Jonathan M.; Myers, Martin G.; Araki, Eichii; White, Morris F.

In: Biochemistry, Vol. 31, No. 37, 1992, p. 9031-9039.

Research output: Contribution to journalArticle

Miralpeix, M, Sun, XJ, Backer, JM, Myers, MG, Araki, E & White, MF 1992, 'Insulin stimulates tyrosine phosphorylation of multiple high molecular weight substrates in Fao hepatoma cells', Biochemistry, vol. 31, no. 37, pp. 9031-9039.
Miralpeix, Montserrat ; Sun, Xiao Jian ; Backer, Jonathan M. ; Myers, Martin G. ; Araki, Eichii ; White, Morris F. / Insulin stimulates tyrosine phosphorylation of multiple high molecular weight substrates in Fao hepatoma cells. In: Biochemistry. 1992 ; Vol. 31, No. 37. pp. 9031-9039.
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