Selective backbone labeling of proteins using 1,2-13C2-pyruvate as carbon source.
A simple isotope labeling approach for selective (13)C/(15)N backbone labeling of proteins is described. Using {1,2-(13)C(2)}-pyruvate as the sole carbon source in bacterial growth media, selective incorporation of (13)C(alpha)-(13)CO spin-pairs into the backbones of protein molecules with medium-to-high levels of (13)C-enrichment is possible for a subset of 12 amino acids. The isotope labeling scheme has been tested on a pair of proteins--a 7-kDa immunoglobulin binding domain B1 of streptococcal protein G and an 82-kDa enzyme malate synthase G. A number of protein NMR applications are expected to benefit from the {1,2-(13)C(2)}-pyruvate based protein production.
Guo
Chenyun
C
Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA.
Geng
Chun
C
Tugarinov
Vitali
V
D001426:Bacterial Proteins / Q000737:chemistry; D002247:Carbon Isotopes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008292:Malate Synthase / Q000737:chemistry; D008954:Models, Biological; D011506:Proteins / Q000737:chemistry*; D019289:Pyruvic Acid / Q000737:chemistry*
D001426:Bacterial Proteins; D002247:Carbon Isotopes; C034284:IgG Fc-binding protein, Streptococcus; D011506:Proteins; D019289:Pyruvic Acid; D008292:Malate Synthase