プロトン・ポンプの高次細胞機能の発現と疾病に関する研究

• Project/Area Number: 15687004
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Functional biochemistry
• Research Institution: Doshisha Women's College of Liberal Arts (2005); Osaka University (2003-2004)
• Principal Investigator: 和田 戈虹 (孫 戈虹) 同志社女子大学, 薬学部, 助教授 (00314427)
• Co-Investigator(Kenkyū-buntansha): 田畑 裕幸 同志社女子大学, 薬学部, 助手 (70378785) ; 川村 暢幸 同志社女子大学, 薬学部, 助手 (30411086)
• Project Period (FY): 2003 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥23,270,000 (Direct Cost: ¥17,900,000、Indirect Cost: ¥5,370,000); Fiscal Year 2005: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2004: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2003: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000)
• Keywords: V-ATPase / 破骨細胞 / endocytosis / renal acidosis / 酸性環境 / 液胞型ATPase / サブユニット・イソフォーム / 組織特異性 / 遺伝子破壊マウス / 液胞型プロトンATPase / 腎アシドーシス / シナプス小胞 / アクロソーム

Research Abstract

動物個体には多彩な酸性環境(オルガネラ、コンパートメント)が存在している。細胞内においては、細胞内膜系のオルガネラ、すなわち、ゴルジ体、エンドソーム、リソソームなどの内部は酸性に保たれている。個体レベルでは、骨吸収窩、尿細管、膀胱、輸精管などの内腔も酸性である。細胞の内外に形成されている酸性環境は、タンパク質分解、小胞輸送をはじめ、免疫・神経機能、骨代謝、イオン恒常維持などに重要な役割を果たしている。酸性環境の異常は骨粗鬆症、大理石病、ガン転移、感音性難聴、腎アシドーシス、不妊症など様々な疾病と密接に関連している。今年度は、腎臓および内分泌における液胞型ATPase(V-ATPase)の生理機能を中心に解析を行った。私たちはサブユニット・イソフォームによってV-ATPaseが細胞あるいは組織特異的な局在を示すことを明らかにした.同時に,特定なオルガネラ膜に特異的な膜タンパク質を複数同定してきた.さらに,リソソーム局在型a3イソフォームが,V-ATPase複合体のまま,細胞形質膜に局在を変化する「リソソームの分泌」と言うべき現象を発見した.これは,極めて興味深い研究成果であり,破骨細胞などの機能を解明する上で重要な知見である.これらの研究成果はNature Cell Biologyをはじめ、複数報の論文に発表した。

Research Products

• [Journal Article] Simple and straightforward construction of a mouse gene targeting vector using in vitro transposon reactions. (2006)
  • Author(s): Aoyama, M., et al.
  • Journal Title: Nucleic Acids Res. 33
• [Journal Article] V-ATPase Interacts with ARNO and Arf6 in Early Endosomes and Regulates the Protein Degradative Pathway. (2006)
  • Author(s): Hurtado-Lorenzo, A., et al.
  • Journal Title: Nat.Cell Biol. 8 Pages: 124-136
• [Journal Article] Selective assembly of subunit isoforms in mouse kidney. (2006)
  • Author(s): Sun-Wada, G-H, et al.
  • Journal Title: J.Bioenerg.Biomemb. (In press)
• [Journal Article] Distinct Expression Patterns of Different Subunit Isoforms of the V-ATPase in the Rat Epididymis. (2005)
  • Author(s): Pietrement, C., et al.
  • Journal Title: Biol Reprod. 74 Pages: 185-194
• [Journal Article] Ubiquitous and Kidney-Specific Subunits of Vacuolar H+-ATPase Are Differentially Expressed during Nephrogenesis. (2005)
  • Author(s): Jouret, F., et al.
  • Journal Title: J.Am.Soc.Nephrol. 16 Pages: 3235-3246
• [Journal Article] Simple and straightforward construction of a mouse gene targeting vector using in vitro transposition reactions (2005)
  • Author(s): Aoyama, M., Agari, K., Sun-Wada, G-H., Futai, M., Wada, Y.
  • Journal Title: Nucleic Acids Research (In press)
• [Journal Article] Diverse and essential roles of mammalian vacuolar-type proton pump ATPase : toward the physiological understanding of inside acidic compartments. (2004)
  • Author(s): Sun-Wada, G-H, Wada, Y., Futai, M.
  • Journal Title: Biochim Biophys Acta. 23号 Pages: 106-114
• [Journal Article] Proton pumping ATPases and diverse inside-acidic compartments. (2004)
  • Author(s): Futai, M., Sun-Wada, G-H, Wada, Y.
  • Journal Title: Yakugaku Zasshi 124 Pages: 243-260
• [Book] ATPase Handbook, Ion Pumping ATPases : Biochemistry, Cell Biology, and Pathophysiology.(eds.Futai, M., Kaplan, J., Wada, Y.) (2004)
  • Author(s): Sun-Wada, G-H, Wada, Y., Futai, M.
  • Publisher: STM Books, Wiley VCH, Weinheim, Germany
• [Book] ATPase Handbook, Ion Pumping ATPases : Biochemistry, Cell Biology, and Pathophysiology.(eds.Futai, M., Kaplan, J., Wada, Y.) (2004)
  • Author(s): Futai, M., Sun-Wada, G-H, Wada, Y.
  • Publisher: STM Books, Wiley VCH, Weinheim, Germany
• [Publications] Sun-Wada, G-H, Kamei, Y., Wada, Y., Futai, M.: "Regulatory elements directing gut-specific expression of GATA6 gene during mouse early development."J.Biochem.. in press. (2004)
• [Publications] Sun-Wada, G-H, Y., Murata, Y., Namba, M., Wada, Y., Futai, M.: "Mouse proton pump ATPase C subunit isoforms (C2-a and C2-b) specifically expressed in kidney and lung."J.Biol.Chem.. 278. 44843-44851 (2003)
• [Publications] Toyomura, T., Murata, Y., Yamamoto, A., Oka, T., Sun-Wada, G-H., Wada, Y., Futai, M.: "From lysosomes to plasma membrane : localization of vacuolar type H^+-ATPase with the a3 isoform during osteoclast differentiation."J.Biol.Chem.. 278. 22023-22030 (2003)
• [Publications] Hirata, T., Iwamoto-Kihara, A., Sun-Wada, G-H, Okajima, T., Wada, Y., Futai, M.: "Subunit rotation of vacuolar-type proton pumping ATPase : Relative rotation of G as to c subunit."J.Biol.Chem.. 278. 23714-23719 (2003)
• [Publications] Sun-Wada, G-H, Yoshimizu, T., Imai-Senga, Y., Wada, Y., Futai, M.: "Diversity of mouse proton-translocating ATPase : Presence of multiple isoforms of the C, d and G subunits."Gene. 302. 147-153 (2003)
• [Publications] Sun-Wada, G-H, Wada, Y., Futai, M.: "Lysosome and lysosome-related organelles responsible for specialized functions in higher organisms."Cell Struct.Func.. 28. 28:455-28:463 (2003)
• [Publications] Sun-Wada, G-H, Wada, Y., Futai, M.: "Vacuolar H^+ pumping ATPases in luminal acidic organelles and extracellular compartments : Common rotational mechanism and diverse physiological roles."J.Bioenerg.Biomemb.. 35. 347-358 (2003)
• [Publications] Sun-Wada, G-H, Wada, Y., Futai, M.: "ATPase Handbook, Ion Pumping ATPases : Biochemistry, Cell Biology, and Pathophysiology.eds.Futai, M., Kaplan, J., Wada, Y."STM Books, Wiley VCH, Weinheim, Germany(in press). (2004)
• [Publications] Futai, M., Sun-Wada, G-H, Wada, Y.: "ATPase Handbook, Ion Pumping ATPases : Biochemistry, Cell Biology, and Pathophysiology.eds.Futai, M., Kaplan, J., Wada, Y."STM Books, Wiley VCH, Weinheim, Germany(in press). (2004)