TY - CHAP
T1 - Selective isotope labeling of recombinant proteins in escherichia coli
AU - Tong, Kit I.
AU - Yamamoto, Masayuki
AU - Tanaka, Toshiyuki
PY - 2012
Y1 - 2012
N2 - Selective stable-isotope labeling is a useful technique to study structures of proteins, especially intrinsically disordered proteins, by nuclear magnetic resonance spectroscopy. Here, we describe a simple method for amino acid selective isotope labeling of recombinant proteins in E. coli. This method only requires addition of an excess of unlabeled amino acids and, if necessary, enzyme inhibitors to the culture medium. Its efficiency has been demonstrated even in labeling with glutamine or glutamate that is easily converted to other amino acid types by the metabolic pathways of E. coli.
AB - Selective stable-isotope labeling is a useful technique to study structures of proteins, especially intrinsically disordered proteins, by nuclear magnetic resonance spectroscopy. Here, we describe a simple method for amino acid selective isotope labeling of recombinant proteins in E. coli. This method only requires addition of an excess of unlabeled amino acids and, if necessary, enzyme inhibitors to the culture medium. Its efficiency has been demonstrated even in labeling with glutamine or glutamate that is easily converted to other amino acid types by the metabolic pathways of E. coli.
KW - Amino acid selective isotope labeling
KW - Intrinsically disordered protein
KW - N-Glutamate labeling
KW - N-Glutamine labeling
UR - http://www.scopus.com/inward/record.url?scp=84865345248&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84865345248&partnerID=8YFLogxK
U2 - 10.1007/978-1-4614-3704-8_30
DO - 10.1007/978-1-4614-3704-8_30
M3 - Chapter
C2 - 22821543
AN - SCOPUS:84865345248
SN - 9781461437031
T3 - Methods in Molecular Biology
SP - 439
EP - 448
BT - Intrinsically Disordered Protein Analysis
A2 - Uversky, Vladimir
A2 - Uversky, Vladimir
A2 - Dunker, Keith
ER -