Methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alabeta and Ilegamma2 methyl groups in large proteins.
A set of sensitive methyl-detected 'out-and-back' NMR experiments for simultaneous assignments of Alabeta and Ilegamma2 methyl positions in large proteins is described. The developed methodology is applied to an 82-kDa enzyme Malate Synthase G. Complete alanine beta and isoleucine gamma2 1H-13C methyl chemical shift assignments could be obtained from the set of new methyl-detected 'out-and-back' 3D experiments. The described methodology for methyl assignments should be applicable to protein molecules within approximately 100-kDa molecular weight range irrespective of the labeling strategy chosen to produce selectively protonated Alabeta and Ilegamma2 13CH3 sites on a deuterated background.
Sheppard
Devon
D
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Guo
Chenyun
C
Tugarinov
Vitali
V
D000409:Alanine / Q000737:chemistry* / Q000378:metabolism; D000465:Algorithms; D002247:Carbon Isotopes / Q000737:chemistry* / Q000378:metabolism; D003198:Computer Simulation; D004926:Escherichia coli / Q000235:genetics; D007532:Isoleucine / Q000737:chemistry* / Q000378:metabolism; D008292:Malate Synthase / Q000737:chemistry; D008745:Methylation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D019289:Pyruvic Acid / Q000378:metabolism; D014633:Valine / Q000737:chemistry / Q000378:metabolism
D002247:Carbon Isotopes; D011506:Proteins; D007532:Isoleucine; D019289:Pyruvic Acid; D008292:Malate Synthase; D014633:Valine; D000409:Alanine