NMR analysis of the structural flexibility of membrane proteins

2015 Fiscal Year Final Research Report

• Project/Area Number: 26891016
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Biophysics
• Research Institution: Osaka 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
• Keywords: NMR / ナノディスク / 無細胞蛋白質合成系 / 膜蛋白質

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.

Free Research Field

蛋白質科学