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
Source: Journal of Biomolecular NMR