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Cell‐free synthesis of 15N‐labeled proteins for NMR studies
Author(s) -
Ozawa Kiyoshi,
Dixon Nicholas E.,
Otting Gottfried
Publication year - 2005
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216540500217859
Subject(s) - cell free protein synthesis , protein biosynthesis , escherichia coli , nuclear magnetic resonance spectroscopy , chemistry , amino acid , in vitro , biochemistry , cell free system , cell , stereochemistry , gene
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.IUBMB Life, 57: 615‐622, 2005