A simple strategy for C-13,H-1 labeling at the Ile-gamma 2 methyl position in highly deuterated proteins

作者：Ruschak Amy M; Velyvis Algirdas; Kay Lewis E^*

来源：Journal of Biomolecular NMR, 2010, 48(3): 129-135.

DOI：10.1007/s10858-010-9449-1

• Methyl labeling
• Ile-gamma 2 methyl
• Supra-molecular complexes
• Proteasome
• HMQC
• Methyl-TROSY

A straightforward approach for the production of highly deuterated proteins labeled with C-13 and H-1 at Ile-gamma 2 methyl positions is described. The utility of the methodology is illustrated with an application involving the half proteasome (360 kDa). High quality 2D Ile C-13(gamma 2),H-1(gamma 2) HMQC data sets, exploiting the methyl-TROSY principle, are recorded with excellent sensitivity and resolution, that compare favorably with Ile C-13(delta 1),H-1(delta 1) spectra. This labeling scheme adds to a growing list of different approaches that are significantly impacting the utility of solution NMR spectroscopy in studies of supra-molecular systems.

• 出版日期2010-11