TECHNOLOGY OF NMR SIGNAL ASSIGNMENTS OF PROTEINS WITH HIGH SENSITIVITY

Research Project

Project/Area Number	23370056
Research Category	Grant-in-Aid for Scientific Research (B)
Allocation Type	Single-year Grants
Section	一般
Research Field	Structural biochemistry
Research Institution	Kitasato University
Principal Investigator	KOHNO TOSHIYUKI 北里大学, 医学部, 講師 (40416657)
Project Period (FY)	2011-04-01 – 2014-03-31
Project Status	Completed (Fiscal Year 2013)
Budget Amount *help	¥20,020,000 (Direct Cost: ¥15,400,000、Indirect Cost: ¥4,620,000) Fiscal Year 2013: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000) Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000) Fiscal Year 2011: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Keywords	生体高分子 / NMR / タンパク質 / 安定同位体 / 微量解析 / 無細胞タンパク質合成 / 安定同位体標識
Research Abstract	We have improved and/or developed two methods for NMR signal assignments of large proteins with less amount of sample. At first, we improved NMR pulse programs for MAGICAL method that we previously developed with the intelligent stable isotope labeling techniques. As a result, the sensitivity of NMR measurements was dramatically enhanced by 30%. Second, we developed a new method for site-directed stable isotope labeling by changing two kinds of tRNAs of major codon and minor codon for the same amino acid. With these methods, it was indicated that NMR signals of proteins could be assigned which were difficult to analyze by using traditional NMR techniques.