Selective 1H-13C NMR spectroscopy of methyl groups in residually protonated samples of large proteins
 

Selective 1H-13C NMR spectroscopy of methyl groups in residually protonated samples of large proteins


Abstract Methyl 13CHD2 isotopomers of all methyl-containing amino-acids can be observed in residually protonated samples of large proteins obtained from [U-13C,1H]-glucose/D2O-based bacterial media, with sensitivity sufficient for a number of NMR applications. Selective detection of some subsets of methyl groups (Alaβ, Thrγ2) is possible using simple â??out-and-backâ?? NMR methodology. Such selective methyl-detected â??out-and-backâ?? NMR experiments allow complete assignments of threonine γ2 methyls in residually protonated, [U-13C,1H]-glucose/D2O-derived samples of an 82-kDa enzyme Malate Synthase G. [U-13C,1H]-glucose/D2O-derived protein samples are relatively inexpensive and are usually available at very early stages of any NMR study of high-molecular-weight systems.

• Content Type Journal Article
• Pages 127-133
• DOI 10.1007/s10858-009-9393-0
• Authors
  • Chenyun Guo, University of Maryland Department of Chemistry and Biochemistry College Park MD 20742 USA
  • Vitali Tugarinov, University of Maryland Department of Chemistry and Biochemistry College Park MD 20742 USA

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738
  • Journal Volume Volume 46
  • Journal Issue Volume 46, Number 2

Source: Journal of Biomolecular NMR