Production of protein for nuclear magnetic resonance study using the wheat germ cell-free system.
Nuclear magnetic resonance (NMR) methods have been developed to determine the three-dimensional structures of proteins, to estimate protein folding, and to discover high-affinity ligands for proteins. However, one of the difficulties encountered in the application of such NMR methods to proteins is that we should obtain milligram quantities of 15N and/or 13C-labeled pure proteins of interest. Here, we describe the method to produce proteins for NMR experiments using the improved wheat germ cell-free system, which exhibits several attractive features for high-throughput NMR study of proteins.
Kohno
Toshiyuki
T
Molecular Structure Research Group, Mitsubishi Kagaku Institute of Life Sciences (MITILS), Tokyo, Japan.
Endo
Yaeta
Y
D002474:Cell-Free System; D007553:Isotope Labeling; D008967:Molecular Biology / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014908:Triticum / Q000196:embryology* / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism
D010940:Plant Proteins; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin