Strategic evaluation of Receptor interaction using resonance energy transfer

• Project/Area Number: 17590058
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Kyoto University
• Principal Investigator: KOSHIMIZU Takaaki Kyoto Uni. Dep. Pharmaceutical Sci, Lecturer, 薬学研究科, 講師 (20392491)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
• Keywords: Receptor Interaction / Drug discovery / Pharmacology / 受容体オリゴマー / BRET / 免疫沈降 / G蛋白質受容体 / ノックアウトマウス / 蛍光蛋白質 / 発光蛋白質 / 受容体薬理学 / 受容体ダイマー / Gタンパク質共役型受容体 / 共鳴エネルギー移動 / 蛍光タンパク質 / 発光タンパク質

Research Abstract

Heteromultimerization of G-protein coupled receptors makes new class of drug targets. We evaluated systemic strategy to delineate functional properties of heteromer receptor, from their molecular level to whole animal level. Combinations of drug-oriented pharmacological method and genetic manipulation of receptor expression levels were equally effective for this purpose.
To elucidate temporal and spacial receptor-receptor interactions in living cells, N-or C-terminus of the subunits were connected with either luciferase or green fluorescent proteins and transfer of bioluminescent resonance energy from luciferase to fluorescent proteins was monitored.
Using gene targeting, physical and functional interactions of α 1-adrenergic receptor subtypes and V1 vasopressin subtypes were investigated. Elucidation of the physiological and pathophysiological roles of the specific α 1-adrenergic receptor subtype has been hampered, because the α 1-adrenergic receptor subtypes are co-expressed in the same arterial smooth muscles with different ratios, and that sufficiently subtype-selective agonists and antagonists have not been available. In addition, recent biochemical and pharmacological studies confirmed the potential role of dimerization of distinct α 1-adrenergic receptor subtypes in controlling their expression and pharmacological properties. Our data showed that α 1B-and α 1D-adrenergic receptors play distinctive contributions to the resting and agonist-stimulated BP regulations, particularly to the progression of hypertensive state. The V1a receptor, on the other hand, plays an important role in normal resting arterial BP regulation mainly by its regulation of circulating blood volume and baroreflex sensitivity.

Research Products

• [Journal Article] Vla Vasopressin receptors maintain normal blood pressure by regulating circulating blood volume and baroreflex sensitivity. (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: Proc Natl Acad Sci USA 103(20)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Carboxy-terminal Splicing Enhances Physical Interactions between the Cytoplasmic Tails of Purinergic P2X Receptors. (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: Mol Pharmacol 69(5) Pages: 1588-1598
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Functional Role of Spliced Cytoplasmic Tails in P2X(2)-Receptor-Mediated Cellular Signaling (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: J Pharmacol Sci 101(4) Pages: 261-266
  • NAID: 130000074469
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Free fatty acids administered into the colon promote the secretion of glucagon-like peptide-1 and insulin. (2006)
  • Author(s): Adachi, T., et al.
  • Journal Title: Biochem Biophys Res Commun 340(1) Pages: 332-337
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] V1a vasopressin receptors maintain normal blood pressure by regulating circulating blood volume and baroreflex sensitivity. (2006)
  • Author(s): Koshimizu, T, Nasa, Y, Tanoue, A, Oikawa, R, Kawahara, Y, Kiyono, Y, Adachi, T, Tanaka, T, Kuwaki, T, Mori, T, Takeo, S, Okamura, H, Tsujimoto, G.
  • Journal Title: Proc Natl Acad Sci USA 103(20) Pages: 7807-12
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Carboxyl-terminal splicing enhances physical interactions between the cytoplasmic tails of purinergic P2X receptors. (2006)
  • Author(s): Koshimizu, T, Kretschmannova, K, He, ML, Ueno, S, Tanoue, A, Yanagihara, N, Stojilkovic, SS, Tsujimoto, G.
  • Journal Title: Mol Pharmacol 69(5) Pages: 1588-98
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Clinical implications from studies of alpha1 adrenergic receptor knockout mice. (2006)
  • Author(s): Koshimizu, T, Tanoue, A, Tsujimoto, G.
  • Journal Title: Biochem Pharmacol. (accepted)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional Role of Spliced Cytoplasmic Tails in P2X(2)-Receptor- Mediated Cellular Signaling. (2006)
  • Author(s): Koshimizu, T, Tsujimoto, G.
  • Journal Title: J Pharmacol Sci 101(4) Pages: 261-6
  • NAID: 130000074469
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Free fatty acids administered into the colon promote the secretion of glucagon-like peptide-1 and insulin. (2006)
  • Author(s): Adachi, T, Tanaka, T, Takemoto, K, Koshimizu, T, Hirasawa, A, Tsujimoto, G.
  • Journal Title: Biochem Biophys Res Commun 340(1) Pages: 332-7
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Vla Vasopressin receptors maintain normal blood pressure by regulating circulating blood volume and baroreflex sensitivity. (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: Proc Natl Acad Sci U S A 103(20) Pages: 1207-1207
• [Journal Article] Vla Vasopressin receptors maintain normal blood pressure by regulating circulating blood volume and baroreflex sensitivity. (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: Proc Natl Acad Sci USA (印刷中)
• [Journal Article] Carboxy-terminal Splicing Enhances Physical Interactions between the Cytoplasmic Tails of Purinergic P2X Receptors. (2006)
  • Author(s): Koshimizu T., et al.
  • Journal Title: Mol Pharmacol (印刷中)
• [Journal Article] Two alphal-adrenergic receptor subtypes regulating the vasopressor response have differential roles in blood pressure regulation. (2005)
  • Author(s): Hosoda, C., Koshimizu T.et al.
  • Journal Title: Mol Pharmacol 67(3) Pages: 912-922
• [Journal Article] Inhibition of protein kinase CK2 prevents the progression of glomerulonephritis. (2005)
  • Author(s): Yamada, M., et al.
  • Journal Title: Proc Natl Acad Sci USA 102(21) Pages: 7736-7741
• [Journal Article] Molecular Dissection of Purinergic P2X Receptor-Channels. (2005)
  • Author(s): Stojilkovic, S.S., et al.
  • Journal Title: Ann NY Acad Sci 1048 Pages: 1-15
• [Journal Article] Clinical implications from studies of alphal adrenergic receptor knockout mice
  • Author(s): Koshimizu T., et al.
  • Journal Title: Biochem Pharmacol (accepted)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Clinical implications from studies of alphal adrenergic receptor knockout mice
  • Author(s): Koshimizu T., et al.
  • Journal Title: Biochem Pharmacol (accepted)