Selective backbone labeling of proteins using {1,2-13C2}-pyruvate as carbon source |
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Authors: | Chenyun Guo Chun Geng Vitali Tugarinov |
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Affiliation: | (1) Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA |
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Abstract: | A simple isotope labeling approach for selective 13C/15N backbone labeling of proteins is described. Using {1,2-13C2}-pyruvate as the sole carbon source in bacterial growth media, selective incorporation of 13Cα-13CO spin-pairs into the backbones of protein molecules with medium-to-high levels of 13C-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 {1,2-13C2}-pyruvate based protein production. |
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Keywords: | Selective isotope labeling {1,2-13C2}-Pyruvate Protein backbone MSG |
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