Structural and functional analysis of tripartite ABC transporter MacB

• Project/Area Number: 18K06079
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Okada Ui 東京工業大学, 生命理工学院, 助教 (20507181)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 構造生物化学 / X線結晶構造解析 / ABC輸送体 / 膜蛋白質 / 薬剤耐性 / 構造生物学 / 輸送タンパク質 / X線結晶解析 / 膜輸送体

Outline of Final Research Achievements

The macrolide efflux transporter MacB is a member of ATP-binding cassette (ABC) transporter which forms a tripartite efflux complex with a membrane fusion protein MacA and an outer membrane channel TolC. The molecular mechanism of the substrate translocation by MacAB-TolC complex remains largely unknown. We determined the first crystal structure of MacB from Acinetobacter baumannii at 3.4 angstrom resolution in 2017. In this study, we have determined the higher resolution structure of MacB homolog at 2.0 angstrom resolution, and several structures from other conditions have also been solved. The structural comparison between these structures revealed the conformational change of MacB, and these results provide new insights into the mechanism of the substrate translocation.

Academic Significance and Societal Importance of the Research Achievements

本研究の対象であるMacBは、マクロライド系抗生物質やペプチド毒素、リポ多糖等の生物学的に重要な物質の排出に関与しており、その排出機構を解明することは、創薬のみならず生命現象の理解という観点からも非常に有用な情報が引き出せると期待される。本研究で明らかとなったMacBの構造変化は、従来のABC輸送体とは大きく異なるものであった。これらの結果から、詳細が解明されていないMacBを含むVII型ABC輸送体の基質輸送の分子機構について、新たなモデルを提唱した。

Research Products

• [Journal Article] Structure and function relationship of OqxB efflux pump from Klebsiella pneumoniae (2021)
  • Author(s): Bharatham Nagakumar、Bhowmik Purnendu、Aoki Maho、Okada Ui、Sharma Sreevalli、Yamashita Eiki、Shanbhag Anirudh P.、Rajagopal Sreenath、Thomas Teby、Sarma Maitrayee、Narjari Riya、Nagaraj Savitha、Ramachandran Vasanthi、Katagihallimath Nainesh、Datta Santanu、Murakami Satoshi
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 1-12
  • DOI: 10.1038/s41467-021-25679-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 三者複合体形成型ABCトランスポーターMacBの構造解析 (2020)
  • Author(s): 岡田 有意、村上 聡
  • Journal Title: Farumashia Volume: 56 Issue: 6 Pages: 499-503
  • DOI: 10.14894/faruawpsj.56.6_499
  • NAID: 130007850168
  • ISSN: 0014-8601, 2189-7026
  • Peer Reviewed
• [Journal Article] Tripartite transporters as mechanotransmitters in periplasmic alternating-access mechanisms. (2020)
  • Author(s): Murakami, Okada, van Veen
  • Journal Title: FEBS Letters Volume: 594 Issue: 23 Pages: 3908-3919
  • DOI: 10.1002/1873-3468.13929
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BpeB, a major resistance-nodulation-cell division transporter from Burkholderia cenocepacia: construct design, crystallization and preliminary structural analysis (2018)
  • Author(s): T. Horikawa, L.-W. Hung, H.-B. Kim, D. Shaya, C.-Y. Kim, T. C. Terwilliger, E. Yamashita, M. Aoki, U. Okada and S. Murakami
  • Journal Title: Acta Crystallogr. F Volume: 74 Issue: 11 Pages: 710-716
  • DOI: 10.1107/s2053230x18013547
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Crystal structure of tripartite-type ABC transporter MacB from Acinetobacter baumannii (2021)
  • Author(s): Okada Ui
  • Organizer: PDB50 Anniversary Symposium in Asia
  • Int'l Joint Research
• [Presentation] Crystal structure of tripartite-type ABC transporter MacB from Acinetobacter baumannii (2019)
  • Author(s): Ui Okada, Eiki Yamashita, Arthur Neuberger, Mayu Morimoto, Hendrik W. van Veen & Satoshi Murakami
  • Organizer: Gordon Research Conference on Multi-Drug Efflux Systems
  • Int'l Joint Research