IRBIT interacts with the catalytic core of phosphatidylinositol phosphate kinase type Ia and IIa through conserved catalytic aspartate residues

Hideaki Ando, Matsumi Hirose, Laura Gainche, Katsuhiro Kawaai, Benjamin Bonneau, Takeshi Ijuin, Toshiki Itoh, Tadaomi Takenawa, Katsuhiko Mikoshiba

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Phosphatidylinositol phosphate kinases (PIPKs) are lipid kinases that generate phosphatidylinositol 4,5-bisphosphate (PI(4,5)P 2 ), a critical lipid signaling molecule that regulates diverse cellular functions, including the activities of membrane channels and transporters. IRBIT (IP 3 R-binding protein released with inositol 1,4,5-trisphosphate) is a multifunctional protein that regulates diverse target proteins. Here, we report that IRBIT forms signaling complexes with members of the PIPK family. IRBIT bound to all PIPK isoforms in heterologous expression systems and specifically interacted with PIPK type Ia (PIPKIa) and type IIa (PIPKIIa) in mouse cerebellum. Site-directed mutagenesis revealed that two conserved catalytic aspartate residues of PIPKIa and PIPKIIa are involved in the interaction with IRBIT. Furthermore, phosphatidylinositol 4-phosphate, Mg 2+ , and/or ATP interfered with the interaction, suggesting that IRBIT interacts with catalytic cores of PIPKs. Mutations of phosphorylation sites in the serine-rich region of IRBIT affected the selectivity of its interaction with PIPKIa and PIPKIIa. The structural flexibility of the serine-rich region, located in the intrinsically disordered protein region, is assumed to underlie the mechanism of this interaction. Furthermore, in vitro binding experiments and immunocytochemistry suggest that IRBIT and PIPKIa interact with the Na + /HCO 3 - cotransporter NBCe1-B. These results suggest that IRBIT forms signaling complexes with PIPKIa and NBCe1-B, whose activity is regulated by PI(4,5)P 2 .

Original languageEnglish
Article number0141569
JournalPloS one
Volume10
Issue number10
DOIs
Publication statusPublished - 2015 Oct 28
Externally publishedYes

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Phosphatidylinositol Phosphates
phosphatidylinositols
aspartic acid
Aspartic Acid
Catalytic Domain
phosphotransferases (kinases)
Phosphotransferases
phosphates
Serine
serine
Intrinsically Disordered Proteins
Lipids
1-phosphatidylinositol 4-kinase
Mutagenesis
Phosphorylation
Inositol 1,4,5-Trisphosphate
Membrane Transport Proteins
Site-Directed Mutagenesis
Ion Channels
proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

IRBIT interacts with the catalytic core of phosphatidylinositol phosphate kinase type Ia and IIa through conserved catalytic aspartate residues. / Ando, Hideaki; Hirose, Matsumi; Gainche, Laura; Kawaai, Katsuhiro; Bonneau, Benjamin; Ijuin, Takeshi; Itoh, Toshiki; Takenawa, Tadaomi; Mikoshiba, Katsuhiko.

In: PloS one, Vol. 10, No. 10, 0141569, 28.10.2015.

Research output: Contribution to journalArticle

Ando, H, Hirose, M, Gainche, L, Kawaai, K, Bonneau, B, Ijuin, T, Itoh, T, Takenawa, T & Mikoshiba, K 2015, 'IRBIT interacts with the catalytic core of phosphatidylinositol phosphate kinase type Ia and IIa through conserved catalytic aspartate residues', PloS one, vol. 10, no. 10, 0141569. https://doi.org/10.1371/journal.pone.0141569
Ando, Hideaki ; Hirose, Matsumi ; Gainche, Laura ; Kawaai, Katsuhiro ; Bonneau, Benjamin ; Ijuin, Takeshi ; Itoh, Toshiki ; Takenawa, Tadaomi ; Mikoshiba, Katsuhiko. / IRBIT interacts with the catalytic core of phosphatidylinositol phosphate kinase type Ia and IIa through conserved catalytic aspartate residues. In: PloS one. 2015 ; Vol. 10, No. 10.
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AU - Bonneau, Benjamin

AU - Ijuin, Takeshi

AU - Itoh, Toshiki

AU - Takenawa, Tadaomi

AU - Mikoshiba, Katsuhiko

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