Production of proteins for NMR studies using the wheat germ cell-free system.
This chapter describes protocols for preparing 15N-labeled proteins (ubiquitin is used as an example) using Escherichia coli cells (with purification) and the wheat germ cell-free system (without purification). A comparison of 1H-15N heteronuclear single-quantum coherence (HSQC) spectra of yeast ubiquitin prepared using each method indicates that this wheat germ cell-free system may be used for rapid nuclear magnetic resonance analyses of proteins without purification.
Kohno
Toshiyuki
T
Laboratory of Structural Biology, Mitsubishi Kagaku Institute of Life Sciences (MITILS), Tokyo, Japan.
D002474:Cell-Free System; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010957:Plasmids; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014908:Triticum / Q000196:embryology* / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism
D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin