PMID:18762866
| Citation |
Tong, KI, Yamamoto, M and Tanaka, T (2008) A simple method for amino acid selective isotope labeling of recombinant proteins in E. coli. J. Biomol. NMR 42:59-67 |
|---|---|
| Abstract |
A simple and user-friendly method of labeling protein selectively with amino acids in vivo is introduced. This technique does not require the use of transaminase-deficient or auxotrophic strains. By manipulating the product feedback inhibitory loops of the E. coli amino acid metabolic pathways and, if necessary, by using enzyme inhibitors, proteins were labeled efficiently in vivo even with amino acid types that are central to the metabolic pathways, such as glutamine. The sequential backbone resonance assignment of the Neh2 domain of Nrf2 transcriptional factor, an intrinsically disordered protein with high spectral degeneracy, was achieved using this labeling method. |
| Links |
PubMed Online version:10.1007/s10858-008-9264-0 |
| Keywords |
Amino Acid Sequence; Amino Acids/analysis; Amino Acids/chemistry; Escherichia coli/chemistry; Escherichia coli/genetics; Escherichia coli/metabolism; Isotope Labeling/methods; Nuclear Magnetic Resonance, Biomolecular; Recombinant Proteins/analysis; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/metabolism |
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