Use of biosynthetic fractional 13C-labeling for backbone NMR assignment of proteins.
We describe a simple approach to classify amino acid residue types in NMR spectra of proteins for supporting the backbone resonance assignments. It makes use of the differences in biosynthetic pathways of the 20 amino acids in Escherichia coli. Therefore, it is distinct from the parameters routinely exploited in the backbone resonance assignment such as chemical shifts and spin topology information. The combination of biosynthetically directed fractional 13C-labeling and uniform 15N-labeling enables us to obtain both residue-type specific information and sequential connectivities from a single protein sample. The residue-type classification exploiting biosynthetic pathways can be used for accelerating the conventional backbone assignment procedure.
Iwai
Hideo
H
Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki, 00014, Finland. hideo.iwai@helsinki.fi
Fiaux
Jocelyne
J
D000596:Amino Acids / Q000737:chemistry; D002247:Carbon Isotopes / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D005947:Glucose / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D015394:Molecular Structure; D009587:Nitrogen Isotopes / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
D000596:Amino Acids; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D005947:Glucose