Cell-free synthesis of 15N-labeled proteins for NMR studies.
Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15 N-labeled proteins from 15 N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR spectra recorded of the crude reaction mixture without concentration or chromatographic purification of the protein. We review methodological aspects of cell-free in vitro protein synthesis based on an Escherichia coli cell extract, in particular with regard to the production of 15 N-labeled proteins for analysis by NMR spectroscopy.
Ozawa
Kiyoshi
K
Research School of Chemistry, Australian National University, Canberra ACT, Australia.
Dixon
Nicholas E
NE
Otting
Gottfried
G
D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D002457:Cell Extracts / Q000191:economics; D002474:Cell-Free System; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000032:analysis / Q000096:biosynthesis / Q000737:chemistry; D007553:Isotope Labeling; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011506:Proteins / Q000378:metabolism*
D000596:Amino Acids; D002457:Cell Extracts; D029968:Escherichia coli Proteins; D009587:Nitrogen Isotopes; D011506:Proteins