Study for mechanism of ABC transporters based on single molecular direct observations using high speed Atomic Force Microscopy.

2016 Fiscal Year Final Research Report

• Project/Area Number: 25293049
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: General physiology
• Research Institution: Keio University
• Principal Investigator: SOHMA Yoshiro 慶應義塾大学, 医学部, 准教授 (60268183)
• Co-Investigator(Renkei-kenkyūsha): UCHIHASHI Takayuki 金沢大学, 数物科学系, 准教授 (30326300) ; NISHIZAKA Takayuki 学習院大学, 理学部, 教授 (40359112) ; SAKURAI Minoru 東京工業大学, バイオ研究基盤支援総合センター, 教授 (50162342) ; SATO Chikara 独立行政法人産業技術総合研究所, バイオメディカル研究部門, 研究グループ長 (00357146)
• Research Collaborator: YAMASHITA Hayato ; YU Ying-Chun ; HWANG Tzyh-Chang
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: ABCトランスポ－タ－ / ATP加水分解 / NBDドメイン / 膜輸送 / 分子間相互作用 / 抗原－抗体反応 / 原子間力顕微鏡

Outline of Final Research Achievements

ABC transporter superfamily is one of the biggest protein families which members play essential physiological roles in human. The members in the ABC transporter superfamily share highly-conserved Nucleotide Binding Domain (NBD) underlying their ATP-driven mechanisms. We investigated molecular mechanism of the ‘NBD engine’ in an ABC member, Cystic Fibrosis Transmembrane Regulator (CFTR) using single-molecular measuring methods including high-speed atomic force microscopy (HS-AFM).
We succeeded the first direct observation of dynamics of the intracellular complex consisting of two NBDs and regulatory (R) domain in CFTR using HS-AFM. We also obtained results in the direct observations of binding/unbinding process between AQP4 and anti-AQP4 antibody as a model of the molecule-molecule interactions. In addition, we performed several molecular physiological/pharmacological studies for disease-associated CFTR mutants.

Free Research Field

分子生理学・薬理学