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Insulin-dependent formation of a complex containing an 85-kDa subunit of phosphatidylinositol 3-kinase and tyrosine-phosphorylated insulin receptor substrate 1.
J Biol Chem 1992; 267(36):25958-65JB

Abstract

Monoclonal antibodies raised against the 85-kDa subunit (p85) of bovine phosphatidylinositol (PI) 3-kinase were found to recognize uncomplexed p85 or p85 in the active PI 3-kinase. Immunoprecipitation studies of Chinese hamster ovary cells, which overexpress the human insulin receptor when treated with insulin, showed increased amounts of p85 and PI 3-kinase activity immunoprecipitable with monoclonal anti-p85 antibody and no increase in the tyrosine phosphorylation of p85. Insulin also induced an association of p85 with the tyrosine-phosphorylated insulin receptor substrate 1 (IRS-1) and other phosphorylated proteins ranging in size from 100 to 170 kDa but not with the activated insulin receptor. In vitro reconstitution studies were used to show p85 in the active PI 3-kinase associated with the tyrosine-phosphorylated IRS-1 but not with the activated insulin receptor. Competition studies using synthetic phosphopeptides corresponding to potential tyrosine phosphorylation sites of IRS-1 revealed that phosphopeptides containing YMXM motifs inhibited this association with different potencies, whereas nonphosphorylated analogues and a phosphopeptide containing the EYYE motif had no effect. Src homology region 2 domains of p85 expressed as glutathione S-transferase fusion proteins also bound to tyrosine-phosphorylated IRS-1. These results suggest that insulin causes the association of PI 3-kinase with IRS-1 via phosphorylated YMXM motifs of IRS-1 and Src homology region 2 domains of p85.

Authors+Show Affiliations

Second Department of Internal Medicine, Kobe University School of Medicine, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

1334490

Citation

Yonezawa, K, et al. "Insulin-dependent Formation of a Complex Containing an 85-kDa Subunit of Phosphatidylinositol 3-kinase and Tyrosine-phosphorylated Insulin Receptor Substrate 1." The Journal of Biological Chemistry, vol. 267, no. 36, 1992, pp. 25958-65.
Yonezawa K, Ueda H, Hara K, et al. Insulin-dependent formation of a complex containing an 85-kDa subunit of phosphatidylinositol 3-kinase and tyrosine-phosphorylated insulin receptor substrate 1. J Biol Chem. 1992;267(36):25958-65.
Yonezawa, K., Ueda, H., Hara, K., Nishida, K., Ando, A., Chavanieu, A., ... Fukui, Y. (1992). Insulin-dependent formation of a complex containing an 85-kDa subunit of phosphatidylinositol 3-kinase and tyrosine-phosphorylated insulin receptor substrate 1. The Journal of Biological Chemistry, 267(36), pp. 25958-65.
Yonezawa K, et al. Insulin-dependent Formation of a Complex Containing an 85-kDa Subunit of Phosphatidylinositol 3-kinase and Tyrosine-phosphorylated Insulin Receptor Substrate 1. J Biol Chem. 1992 Dec 25;267(36):25958-65. PubMed PMID: 1334490.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Insulin-dependent formation of a complex containing an 85-kDa subunit of phosphatidylinositol 3-kinase and tyrosine-phosphorylated insulin receptor substrate 1. A1 - Yonezawa,K, AU - Ueda,H, AU - Hara,K, AU - Nishida,K, AU - Ando,A, AU - Chavanieu,A, AU - Matsuba,H, AU - Shii,K, AU - Yokono,K, AU - Fukui,Y, PY - 1992/12/25/pubmed PY - 1992/12/25/medline PY - 1992/12/25/entrez SP - 25958 EP - 65 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 267 IS - 36 N2 - Monoclonal antibodies raised against the 85-kDa subunit (p85) of bovine phosphatidylinositol (PI) 3-kinase were found to recognize uncomplexed p85 or p85 in the active PI 3-kinase. Immunoprecipitation studies of Chinese hamster ovary cells, which overexpress the human insulin receptor when treated with insulin, showed increased amounts of p85 and PI 3-kinase activity immunoprecipitable with monoclonal anti-p85 antibody and no increase in the tyrosine phosphorylation of p85. Insulin also induced an association of p85 with the tyrosine-phosphorylated insulin receptor substrate 1 (IRS-1) and other phosphorylated proteins ranging in size from 100 to 170 kDa but not with the activated insulin receptor. In vitro reconstitution studies were used to show p85 in the active PI 3-kinase associated with the tyrosine-phosphorylated IRS-1 but not with the activated insulin receptor. Competition studies using synthetic phosphopeptides corresponding to potential tyrosine phosphorylation sites of IRS-1 revealed that phosphopeptides containing YMXM motifs inhibited this association with different potencies, whereas nonphosphorylated analogues and a phosphopeptide containing the EYYE motif had no effect. Src homology region 2 domains of p85 expressed as glutathione S-transferase fusion proteins also bound to tyrosine-phosphorylated IRS-1. These results suggest that insulin causes the association of PI 3-kinase with IRS-1 via phosphorylated YMXM motifs of IRS-1 and Src homology region 2 domains of p85. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/1334490/Insulin_dependent_formation_of_a_complex_containing_an_85_kDa_subunit_of_phosphatidylinositol_3_kinase_and_tyrosine_phosphorylated_insulin_receptor_substrate_1_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=1334490 DB - PRIME DP - Unbound Medicine ER -