A simple method for amino acid selective isotope labeling of recombinant proteins in E. coli.
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.
Tong
Kit I
KI
Center for Tsukuba Advanced Research Alliance, Graduate School of Comprehensive Human Sciences, JST-ERATO Environmental Response Project, University of Tsukuba, Tsukuba, 305-8577, Japan.
Yamamoto
Masayuki
M
Tanaka
Toshiyuki
T