Analysis of NAD metabolic pathway in Mycobacterium tuberculosis and elucidation of its compensatory pathway

• Project/Area Number: 15K07377
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: MORI Shigetarou 国立感染症研究所, 細菌第二部, 室長 (60425676)
• Project Period (FY): 2015-10-21 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 結核菌 / タンパク質 / 酵素 / ドッキングシミュレーション / NAD生合成 / タンパク質発現 / 立体構造解析 / NAD合成経路

Outline of Final Research Achievements

In this research project, we mainly analyzed the functions and structures of mycobacterial enzymes involved in the NAD biosynthetic pathway. The optimal conditions for expressing Rv0212c, Rv1901c, Rv3199c, and Rv3393 of Mycobacterium tuberculosis in Escherichia coli were determined. We found that Rv3393 shows nucleoside hydrolysis activity. In addition, we showed that the structure of the substrate-binding site of phosphoribosyl diphosphate synthase from M. smegmatis differs from that of human phosphoribosyl diphosphate synthase. Furthermore, the enzymatic properties of Rv3257c and Rv3308 from M. tuberculosis and the three-dimensional structure of the nucleotide phosphorylase from M. avium were determined in detail.

Research Products

• [Journal Article] Biochemical and structural investigations on phosphoribosylpyrophosphate synthetase from Mycobacterium smegmatis (2017)
  • Author(s): Donini Stefano、Garavaglia Silvia、Ferraris Davide M.、Miggiano Riccardo、Mori Shigetarou、Shibayama Keigo、Rizzi Menico
  • Journal Title: PLOS ONE Volume: 12 Issue: 4 Pages: e0175815-e0175815
  • DOI: 10.1371/journal.pone.0175815
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Purification and functional characterization of diadenosine 5', 5'''-P1,P4-tetraphosphate phosphorylases from Mycobacterium smegmatis and Mycobacterium avium. (2015)
  • Author(s): Naoko Honda, Hyun Kim, Emiko Rimbara, Atsushi Kato, Keigo Shibayama, Shigetarou Mori
  • Journal Title: Protein Expression and Purification Volume: 112 Pages: 37-42
  • DOI: 10.1016/j.pep.2015.04.010
  • Peer Reviewed / Open Access
• [Presentation] 結核菌由来diadenosine tetraphosphate加リン酸分解酵素の新規阻害剤と抗結核菌活性 (2017)
  • Author(s): 森茂太郎、本田尚子、金玄、林原絵美子、柴山恵吾
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都
  • Year and Date: 2017-03-18
• [Presentation] Molecular characterization of a nicotinate phosphoribosyltransferase from Mycobacterium tuberculosis H37Rv (2017)
  • Author(s): Shigetarou Mori, Hyun Kim, Emiko Rimbara, Keigo Shibayama
  • Organizer: US-Japan Cooperative Medical Science Program: Mycobacterial Panel Meeting
  • Place of Presentation: 韓国・ソウル
  • Year and Date: 2017-02-09
  • Int'l Joint Research
• [Presentation] The development of novel structure based inhibitors of diadenosine phosphorylase from Mycobacterium tuberculosis (2017)
  • Author(s): 森茂太郎, 本田尚子, 金玄, 林原絵美子, 柴山恵吾
  • Organizer: International Union of Microbiological Societies Congresses 2017
  • Int'l Joint Research
• [Presentation] Novel inhibitors of diadenosine tetraphosphate phosphorylase and their antituberculosis activity (2016)
  • Author(s): Shigetarou Mori, Naoko Honda, Hyun Kim, Emiko Rimbara, and Keigo Shibayama
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪市
  • Year and Date: 2016-03-23