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 α - 13 CO spin-pairs into the backbones of protein molecules with medium-to-high...

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Bibliographic Details
Published inJournal of biomolecular NMR Vol. 44; no. 3; pp. 167 - 173
Main Authors Guo, Chenyun, Geng, Chun, Tugarinov, Vitali
Format Journal Article
LanguageEnglish
Published Dordrecht Springer Netherlands 01.07.2009
Springer Nature B.V
Subjects
Bacteria
Biochemistry
Biological and Medical Physics
Biophysics
Communication
Microbiology
Physics
Physics and Astronomy
Spectroscopy/Spectrometry
MSG
Pyruvate
C
Protein backbone
Selective isotope labeling
1,2
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Summary: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 α - 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.
ISSN:0925-2738
1573-5001
DOI:10.1007/s10858-009-9326-y