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A strategy for the introduction of (1H,13C-methyl)-alanine into perdeuterated proteins is described. Specific protonation of alanine methyl groups to a level of 95% can be achieved by overexpressing proteins in M9/D2O based bacterial growth medium supplemented with 800 mg/l of 2-[2H], 3-[13C] l-alanine. However, though simple, this approach results in undesired, non-specific background labeling due...
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