TECHNOLOGY OF NMR ANALYSES FOR A ULTRA LOW AMOUNT OF PROTEIN

• Project/Area Number: 20200035
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)
• Allocation Type: Single-year Grants
• Research Field: Living organism molecular science; Structural biochemistry
• Research Institution: The Institute of Physical and Chemical Research (2010); Mitsubishi Kagaku Institute of Life Sciences (2008-2009)
• Principal Investigator: KOHNO Toshiyuki The Institute of Physical and Chemical Research, NMRパイプライン高度化研究チーム, 客員研究員 (40416657)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥31,460,000 (Direct Cost: ¥24,200,000、Indirect Cost: ¥7,260,000); Fiscal Year 2010: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000); Fiscal Year 2009: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000); Fiscal Year 2008: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
• Keywords: 生物物理 / NMR / 無細胞蛋白質合成 / シグナル帰属 / 選択的安定同位体標識 / 無細胞タンパク質合成 / 安定同位体標識

Research Abstract

We have developed the MAGICAL method for NMR analyses of protein by the combination of the improvement of NMR pulse programs and the improvement of stable isotope labeling techniques. As a result, the sensitivities of NMR measurements are dramatically enhanced and the proteins with low concentration can be analyzed with much less NMR experiment time. We applied to this improved MAGICAL method for less soluble proteins and large proteins. Thus, it was indicated that NMR signals of proteins could be assigned which were difficult to analyze by using traditional NMR techniques.