PTB domain of insulin receptor substrate-1 binds inositol compounds

Hiroshi Takeuchi, Miho Matsuda, Tada Aki Yamamoto, Takashi Kanematsu, Ushio Kikkawa, Hitoshi Yagisawa, Yutaka Watanabe, Masato Hirata

Research output: Contribution to journalArticle

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Abstract

We examined whether a phosphotyrosine binding (PTB) domain from the human insulin receptor substrate-1 (hIRS-1) is capable of binding inositol phosphates/phosphoinositides. The binding specificity was compared with that of the pleckstrin homology (PH) domain derived from the same protein because the three dimensional structure was found to be very similar to that of the PH domain, despite the lack of sequence similarity. We also attempted to locate the site of binding of the inositol compounds. The PTB domain bound [3H]Ins(1,4,5)P3, which was displaced most strongly by Ins(1,3,4,5,6)P5 and InsP6, indicating that these inositol polyphosphates show the highest affinity. The PTB domain bound to liposomes containing PtdIns(4,5)P2, PtdIns(3,4,5)P3 and PtdIns(3,4)P2, but not phosphatidylinositol. In contrast, the PH domain showed a preference for Ins(1,4,5)P3, the polar head of PtdIns(4,5)P2. Site-directed mutagenesis studies were performed to map the binding site for inositol phosphates in the PTB domain. Mutation of K169Q, K171Q or K177Q, located in the loop connecting the β1 and β2 strands, which is partially responsible for binding inositol phosphates/phosphoinositides in the PH domains of several other proteins, reduced binding activity, probably because of a reduction in affinity. Mutation of R212Q or R227Q, shown to be involved in the binding of a phosphotyrosine, had little effect on the binding capacity. These results indicate that the PTB domain of hIRS-1 can bind inositol phosphates/phosphoinositides. Therefore signalling through the PTB domain could be regulated by the binding not only of proteins with phosphotyrosine but also of inositol phosphates/phosphoinositides, implying that PTB domains could be involved in a myriad of interconnections between intracellular signalling pathways.

Original languageEnglish
Pages (from-to)211-218
Number of pages8
JournalBiochemical Journal
Volume334
Issue number1
DOIs
Publication statusPublished - Aug 15 1998

Fingerprint

Insulin Receptor Substrate Proteins
Phosphotyrosine
Inositol
Inositol Phosphates
Phosphatidylinositols
Phosphatidylinositol 4,5-Diphosphate
Binding Sites
Polyphosphates
Mutagenesis
Mutation
Proteins
Site-Directed Mutagenesis
Protein Binding
Liposomes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Takeuchi, H., Matsuda, M., Yamamoto, T. A., Kanematsu, T., Kikkawa, U., Yagisawa, H., ... Hirata, M. (1998). PTB domain of insulin receptor substrate-1 binds inositol compounds. Biochemical Journal, 334(1), 211-218. https://doi.org/10.1042/bj3340211

PTB domain of insulin receptor substrate-1 binds inositol compounds. / Takeuchi, Hiroshi; Matsuda, Miho; Yamamoto, Tada Aki; Kanematsu, Takashi; Kikkawa, Ushio; Yagisawa, Hitoshi; Watanabe, Yutaka; Hirata, Masato.

In: Biochemical Journal, Vol. 334, No. 1, 15.08.1998, p. 211-218.

Research output: Contribution to journalArticle

Takeuchi, H, Matsuda, M, Yamamoto, TA, Kanematsu, T, Kikkawa, U, Yagisawa, H, Watanabe, Y & Hirata, M 1998, 'PTB domain of insulin receptor substrate-1 binds inositol compounds', Biochemical Journal, vol. 334, no. 1, pp. 211-218. https://doi.org/10.1042/bj3340211
Takeuchi, Hiroshi ; Matsuda, Miho ; Yamamoto, Tada Aki ; Kanematsu, Takashi ; Kikkawa, Ushio ; Yagisawa, Hitoshi ; Watanabe, Yutaka ; Hirata, Masato. / PTB domain of insulin receptor substrate-1 binds inositol compounds. In: Biochemical Journal. 1998 ; Vol. 334, No. 1. pp. 211-218.
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