Development of the micro-flow ultra-rapid pulse label NMR apparatus and elucidation of the prion self-replication process.

2016 Fiscal Year Final Research Report

• Project/Area Number: 26291011
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Gifu University
• Principal Investigator: Kuwata Kazuo 岐阜大学, 大学院連合創薬医療情報研究科, 教授 (00170142)
• Co-Investigator(Kenkyū-buntansha): 鎌足 雄司 岐阜大学, 学内共同利用施設等, 助教 (70342772)
• Co-Investigator(Renkei-kenkyūsha): FUKUOKA Mayuko 岐阜大学, 大学院連合創薬医療情報研究科, 特任助教 (60790988)
• Research Collaborator: HONDA Ryo ; Ferdausi Ali ; Kabir Aurangazeb
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: NMR / 混合器 / パルスラベル / プリオン / 自己複製

Outline of Final Research Achievements

To perform the rapid mixing in the NMR magnet and to observe the folding or aggregation reaction of a protein in the atomic resolution, we devised an apparatus for the mixing of two solutions. Denatured protein solution goes through a first tubing and refolding buffer second tubing, and both tubing are connected to a carefully designed mixer in a NMR tube in the magnet. Waste solution is removed from the third tubing thereby enabling the observation in a continuous flow in the NMR tube. We reformed the apparatus used for in cell NMR using the glass material with homogeneous magnetic susceptibility with the aqueous solution. Thus we could remove the false peaks originated from the inhomogeneous magnetic susceptibility that was observed in the previous version of the apparatus. Also we found the ‘A-state’ which corresponds to the branching point between the cellular form and the aggregate, and that the cellular form itself inhibits the aggregate formation uncompetitively.

Free Research Field

構造生物学