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NMR analysis of the structural flexibility of membrane proteins

Research Project

Project/Area Number 26891016
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Biophysics
Research InstitutionOsaka University

Principal Investigator

Ozawa Kiyoshi  大阪大学, たんぱく質研究所, 研究員 (20251770)

Project Period (FY) 2014-08-29 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
KeywordsNMR / ナノディスク / 無細胞蛋白質合成系 / 膜蛋白質 / 蛋白質標識
Outline of Final Research Achievements

pHtrII(1-159) protein is a bacterial phototaxis transducer membrane protein from Natronomonas pharaonis and functions as a cognate transducer to the pharaonis phoborhodpsin. The full length of this transducer has yet to be obtained owing mainly to the difficulties in obtaining this complex in a stable form that eases its structural and functional studies.
In this study, I attempted the use of the phospholipid bilayer nanodisc for studying the structure of pHtrII(1-159) in solution NMR. Cell-free expressed 15N, 2H-labelled pHtrII (1-159) transducer was purified, and it was successfully reconstituted into a nanodisc. Consequently the NMR analysis of the pHtrII (1-159) transducer in a nanodisc provides important information on structural changes between pH6.5 and pH7.0, and between with and without nanodiscs.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Annual Research Report
  • Research Products

    (3 results)

All 2016 2015

All Presentation (1 results) (of which Int'l Joint Research: 1 results) Book (2 results)

  • [Presentation] Cell-free protein synthesis for NMR structural analysis of large proteins and complexes.2015

    • Author(s)
      OzawaKiyoshi
    • Organizer
      Plant and Food Research Meeting
    • Place of Presentation
      オークランド(ニュージーランド)
    • Year and Date
      2015-04-21
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Book] Application of NMR Spectroscopy2016

    • Author(s)
      Kralicek AV and Ozawa K
    • Total Pages
      28
    • Publisher
      Bentham Science Publishers
    • Related Report
      2015 Annual Research Report
  • [Book] Applications of NMR Spectroscopy in Molecular Identification (Structural Biology)2015

    • Author(s)
      Andrew V. Kralicek and Kiyoshi Ozawa
    • Total Pages
      75
    • Publisher
      Bentham Science Publishers
    • Related Report
      2014 Annual Research Report

URL: 

Published: 2014-09-09   Modified: 2017-05-10  

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