Resolution and sensitivity enhancement of heteronuclear correlation for methylene resonances via 2H enrichment and decoupling.

Monodeuterated methylene positions exhibit substantially superior spectral characteristics in 1H−13C correlation experiments as compared to diprotio signals. A combination of 2H decoupling and multiplet editing of HMQC and HSQC experiments provides resolution enhancement for both stereoselective and random fractionally deuterated samples. For HMQC experiments with [2-2Hr, 2-13C]glycine-labeled E. coli thioredoxin (11.7 kDa), 3-fold increases in both 1H and 13C resolution result in a complementary 9-fold enhancement in sensitivity. Owing to a smaller improvement in 13C resolution, the corresponding enhancements for the HSQC experiment are 2-fold less.