Analysis of the voltage dependence of Gq coupled receptors

• Project/Area Number: 26460307
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General physiology
• Research Institution: National Institute for Physiological Sciences
• Principal Investigator: MICHIHIRO TATEYAMA 生理学研究所, 分子細胞生理研究領域, 准教授 (30276472)
• Co-Investigator(Renkei-kenkyūsha): NAKAJO Koichi 大阪医科大学, 医学部, 准教授 (80390699)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2014: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: Gタンパク質共役型受容体 / FRET / 膜電位依存性 / 構造変化 / GPRC / 電位依存性

Outline of Final Research Achievements

It has been reported that the ligand affinities of several G protein coupled receptors are changed depending on the membrane potential. Here we investigated the effects of Gq coupled receptors on the KCNK13 channel current and found that in the purinergic P2Y1R concentration-response relationships were different when the holding potential was different. The voltage dependent difference was abolished when the aspartate residue at the transmembrane domain was mutated to alanine or asparagine. Similar results were obtained from the analysis of FRET between P2Y1R-YFP and CFP-Gq. FRET efficiency was increased by the application of agonist in a concentration dependent manner, and the concentration-FRET curve was leftward shifted by the depolarization of the membrane potential. The shift was not observed when the aspartate residue was mutated to aspargine.

Research Products

• [Journal Article] Stabilizing effects of G protein on the active conformation of adenosine A1 receptor differ depending on G protein type (2016)
  • Author(s): Tateyama M, Kubo Y
  • Journal Title: European Journal of Pharmacology Volume: 788 Pages: 122-131
  • DOI: 10.1016/j.ejphar.2016.06.025
  • Peer Reviewed
• [Journal Article] Auto-oxidation products of epigallo-catechin gallate activate TRPA1 and TRPV1 in sensory neurons. (2015)
  • Author(s): Kurogi, M., Kawai, Y., Nagatomo, K., Tateyama, M., Kubo, Y. and Saitoh, O.
  • Journal Title: Chemical Senses Volume: 40(1) Issue: 1 Pages: 27-46
  • DOI: 10.1093/chemse/bju057
  • Peer Reviewed / Open Access
• [Journal Article] FRET analysis of the activation of GPCR (2014)
  • Author(s): 立山充博、久保義弘
  • Journal Title: Folia Pharmacologica Japonica Volume: 143 Issue: 5 Pages: 249-253
  • DOI: 10.1254/fpj.143.249
  • NAID: 130004827697
  • ISSN: 0015-5691, 1347-8397
• [Presentation] FRET analyses of voltage dependence of P2Y1R (2017)
  • Author(s): 立山充博、久保義弘
  • Organizer: 第94回日本生理学会大会
  • Place of Presentation: アクトシティ浜松（静岡県浜松市）
• [Presentation] Regulation of KCNK13 channel by Gi/o or Gq coupled receptors (2016)
  • Author(s): 立山充博、久保義弘
  • Organizer: 第93回日本生理学会大会
  • Place of Presentation: 北海道・札幌市・札幌コンベンションセンタ－
  • Year and Date: 2016-03-22
• [Presentation] Analyses of the voltage dependence and its structural determinant of the P2Y1 purinergic receptor. (2016)
  • Author(s): 王 培冰、立山充博、久保義弘
  • Organizer: 第93回日本生理学会大会
  • Place of Presentation: 北海道・札幌市・札幌コンベンションセンタ－
  • Year and Date: 2016-03-22
• [Presentation] The Gi/o coupled muscarinic receptors form complex with the G protein gated inwardly rectifying potassium channel. (2015)
  • Author(s): Tateyama M、Kubo Y
  • Organizer: 8th FAOPS
  • Place of Presentation: Bangkok, Thailand
  • Year and Date: 2015-11-23
  • Int'l Joint Research
• [Presentation] The Gi/o coupled muscarinic receptors form complex with the G protein gated inwardly rectifying potassium channel for the efficient channel activation. (2015)
  • Author(s): 立山充博、久保義弘
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 兵庫県・神戸市・神戸国際展示場
  • Year and Date: 2015-07-28
• [Presentation] The Gi/o coupled muscarinic receptors form signaling complex with the G protein dependent inwardly rectifying potassium channe (2015)
  • Author(s): 立山充博、久保義弘
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸国際展示場（兵庫県神戸市）
  • Year and Date: 2015-03-21
• [Presentation] The KCNK13 channel current is increased by the activation of either the Gi/o- or the Gq-coupled receptor. (2015)
  • Author(s): 松原美紀、立山充博、久保義弘
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸国際展示場（兵庫県神戸市）
  • Year and Date: 2015-03-21
• [Presentation] Activation efficiency of the G protein gated inwardly rectifying potassium channel depends on distance from the Gq but not Gi/o coupled receptors (2014)
  • Author(s): 立山充博、久保義弘
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-09-11