Selective labeling strategy, tolerant of amino-acid scrambling, for various expression systems

2016 Fiscal Year Final Research Report

• Project/Area Number: 26650027
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: KASAI Takuma 国立研究開発法人理化学研究所, 生命システム研究センター, 研究員 (70446516)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 蛋白質 / NMR / アミノ酸選択的安定同位体標識 / 符号化標識法 / ピークフィッティング / レプリカ交換モンテカルロ法

Outline of Final Research Achievements

Amino-acid selective stable isotope labeling is a powerful method for signal assignment processes for protein analyses with NMR. This method relies on expression systems with suppressed amino-acid scrambling, such as cell-free protein synthesis. The basic idea of this study is to estimate change of isotope labeling ratio derived from the amino-acid scrambling as unknown parameters from NMR spectra. Such approach requires a method for searching huge parameter space. Therefore we developed a parameter-optimization method for analyses of NMR spectra of amino-acid selectively isotope labeled proteins, using replica exchange Monte Carlo. This method also enabled analyses of low signal-to-noise-ratio spectra.

Free Research Field

構造生物学