Identification of key residues for control of the coupling between ATP hydrolysis and substrate transport of an ABC transporter

• Project/Area Number: 26840048
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Kyoto University
• Principal Investigator: Yamaguchi Tomohiro 京都大学, 薬学研究科(研究院), 助教 (80346791)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 膜タンパク質 / ABCトランスポーター / X線結晶構造解析 / 基質輸送 / ATP

Outline of Final Research Achievements

Human ABC transporters are predicted to have a characteristic mechanism to control the substrate transport driven by hydrolysis of ATP. In this study, structural and functional analyses were performed using CmABCB1 mutant designed based on the crystal structures of CmABCB1, the function of which is similar to that of human ABC transporter. The analyses revealed that glutamine and alanine residues localized on the substrate transport pathway in the transmembrane domain of CmABCB1 act as an important role in the control mechanism.

Research Products

• [Journal Article] Structural and Pharmacological Studies of an ABC Multidrug Transporter (2016)
  • Author(s): 山口知宏
  • Journal Title: YAKUGAKU ZASSHI Volume: 136 Issue: 2 Pages: 197-202
  • DOI: 10.1248/yakushi.15-00229-5
  • NAID: 130005121478
  • ISSN: 0031-6903, 1347-5231
  • Year and Date: 2016-02-01
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of the structural and functional roles of coupling helices in the ATP-binding cassette transporter MsbA through enzyme assays and molecular dynamics simulations (2014)
  • Author(s): Furuta, T., Yamaguchi, T., Kato, H., Sakurai, M.
  • Journal Title: Biochemistry Volume: 53 Issue: 26 Pages: 4261-4272
  • DOI: 10.1021/bi500255j
  • Peer Reviewed
• [Journal Article] Structure and Mechanism of a Eukaryotic ABC Multidrug Transporter (2014)
  • Author(s): 小段篤史、山口知宏、中津亨、加藤博章
  • Journal Title: Nihon Kessho Gakkaishi Volume: 56 Issue: 4 Pages: 224-229
  • DOI: 10.5940/jcrsj.56.224
  • NAID: 130004688353
  • ISSN: 0369-4585, 1884-5576
  • Peer Reviewed
• [Presentation] Serial Femtosecond Crystallography of ABC Transporter (2016)
  • Author(s): Oyama Ryo, Pan Dongqing, Nakatsu Toru, Sato Tomomi, Yamaguchi Tomohiro, Kodan Atsushi, Ueda Kazumitsu, Iwata So, Kato Hiroaki
  • Organizer: 6th Special Meeting, ATP-Binding Cassette (ABC) Proteins: From Multidrug Resistance to Genetic Diseases
  • Place of Presentation: Hotel Grauer Bar (Innsbruck, Austria)
  • Year and Date: 2016-03-05
  • Int'l Joint Research
• [Presentation] Crystallographic study of the ABC transporter CmABCB1 covalently bound to substrate derivatives (2016)
  • Author(s): Matsuoka Keita, Hirokane Ryo, Kodan Atsushi, Yamaguchi Tomohiro, Nakatsu Toru, Ueda Kazumitsu, Kato Hiroaki
  • Organizer: 6th Special Meeting, ATP-Binding Cassette (ABC) Proteins: From Multidrug Resistance to Genetic Diseases
  • Place of Presentation: Hotel Grauer Bar (Innsbruck, Austria)
  • Year and Date: 2016-03-05
  • Int'l Joint Research
• [Presentation] Altering stability of a transmembrane protein, MsbA by structural comparison with its thermophilic homolog (2015)
  • Author(s): Ka Lu, Yamaguchi Tomohiro, Nakatsu Toru, Kato Hiroaki
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015
  • Place of Presentation: Honolulu, Hawaii, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] NMR解析を目指したABCトランスポーターの大腸菌高密度培養による安定同位体標識系の確立 (2015)
  • Author(s): 小田健人、山口知宏、宮ノ入洋平、中津 亨、甲斐荘正恒、加藤博章
  • Organizer: 日本薬学会近畿支部総会・大会
  • Place of Presentation: 大阪大谷大学（大阪府富田林市）
  • Year and Date: 2015-10-17
• [Presentation] ABC多剤排出トランスポーターの構造と機能からADMEを探る (2015)
  • Author(s): 山口知宏
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 神戸学院大学（神戸市）
  • Year and Date: 2015-03-25 – 2015-03-28
  • Invited
• [Presentation] X線自由電子レーザーを用いたタンパク質Ｘ線結晶構造解析 (2015)
  • Author(s): 潘東青、中津亨、山口知宏、小段篤史、小川和浩、松嶌一樹、山下恵太郎、城地保昌、登野健介、矢橋牧名、鈴木守、南後恵理子、岩田想、加藤 博章
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 神戸学院大学（神戸市）
  • Year and Date: 2015-03-25 – 2015-03-28
• [Presentation] Trp変異導入によるABC多剤排出トランスポーターと基質の相互作用解析 (2015)
  • Author(s): 大塚哲央、山口知宏、中津亨、加藤博章
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 神戸学院大学（神戸市）
  • Year and Date: 2015-03-25 – 2015-03-28
• [Presentation] ABC多剤排出トランスポーターの立体構造の特徴 (2014)
  • Author(s): 山口知宏、小段篤史、中津亨、植田和光、加藤博章
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都市）
  • Year and Date: 2014-10-15 – 2014-10-17
  • Invited
• [Presentation] Lysの13C-ジメチル化によるABC多剤排出トランスポーターのNMR解析 (2014)
  • Author(s): 宇都宮裕人、山口知宏、宮ノ入洋平、武田光広、中津亨、甲斐荘正恒、加藤博章
  • Organizer: 第64回日本薬学会近畿支部総会・大会
  • Place of Presentation: 京都薬科大学（京都市）
  • Year and Date: 2014-10-11
• [Presentation] SACLAによるABCトランスポーターおよびホタル・ルシフェラーゼのＸ線結晶構造解析 (2014)
  • Author(s): 潘東青、中津亨、山口知宏、小段篤史、小川和浩、松嶌一樹、城地保昌、登野健介、矢橋牧名、鈴木守、南後恵理子、岩田想、加藤 博章
  • Organizer: 第14回日本蛋白質科学会年会
  • Place of Presentation: ワークピア横浜/横浜産貿ホール マリネリア（横浜市）
  • Year and Date: 2014-06-25 – 2014-06-27