Ultra-fast NMR measurement to analyze dynamics of Sox2 in cells

2014 Fiscal Year Final Research Report

• Project/Area Number: 25650056
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Suntory Foundation for Life Sciences
• Principal Investigator: SUGASE Kenji 公益財団法人サントリー生命科学財団, その他部局等, 主席研究員 (00300822)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: in-cell NMR / タンパク質 / 揺らぎ / 高速測定

Outline of Final Research Achievements

We developed methods that enable rapid measurement of protein dynamics in cells using NMR. Specifically, we found that (i) a 2%-DMSO treatment of cells drastically enhanced the efficiency of protein incorporation into cells using the cell-penetrating peptide, (ii) presaturation of the water signal is useful for rapid NMR measurements to detect flexible regions in a protein, and (iii) the non-uniform sampling (NUS) method was utilized to reduce the total amount of experimental time and the SIFT algorithm, which we have been developing a computer program to reconstruct NUS data, was further developed. As a result, these newly developed methods enabled us to measure an NMR spectrum more rapidly by approximately one-third than the conventional measurement.

Free Research Field

構造生物学