Methyl and Amino Acid Type Labeling
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Selective Isotope-Labeling Methods for Protein Structural Studies
- Selective Isotope-Labeling Methods for Protein Structural Studies
- Stereospecific Leu/Val Methyl Labeling: An Important Technology for NMR Studies of High-Molecular-Weight Complexes (Application Note 48)
- Isotope Labeling of Alanine Methyl Groups on a Deuterated Background for NMR Studies of High-Molecular-Weight Proteins (Application Note 25)
- Production of U-[2H], Thy-γ2[13CH3] Labeled Proteins for methyl-TROSY NMR (Application Note 39)
Methyl labeling refers to methods which introduce either 13CH3 or 13CHD2 groups into otherwise a uniform deuterated protein. This can be achieved in both prokaryotic and eukaryotic expression systems by adding the amino acids(s) (in sufficient amounts) directly to the cell culture medium prior to protein induction. For E. coli cultures, selective protonation on methyl groups of ILV can be achieved by adding alpha-keto acid precursors to the D2O-based cell culture medium 1 hr prior to induction. Alpha-ketoisobutyrate becomes converted to isoleucine, and alpha-ketoisovaleric acid becomes converted to leucine and valine. This methodology has dramatically increased the useful molecular weight range of protein and protein complexes assessable by NMR when using methyl TROSY techniques.
Kevin Millis, PhD
Senior Scientist, Application Development Manager
Kevin Millis, PhD, is the Senior Scientist and Market Development Manager for all NMR and mass spectrometry product lines. Kevin is responsible for Technical Services both internally and externally for all CIL customers as well as being responsible for the application and market development for the CIL products.
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