Incorporating 1H chemical shift determination into 13C-direct detected spectroscopy of intrinsically disordered proteins in solution

Exclusively heteronuclear 13C-detected NMR spectroscopy of proteins in solution has seen resurgence in the past several years. For disordered or unfolded proteins, which tend to have poor 1H-amide chemical shift dispersion, these experiments offer enhanced resolution and the possibility of complete heteronuclear resonance assignment at the cost of leaving the 1H resonances unassigned. Here we report two novel 13C-detected NMR experiments which incorporate a 1H chemical shift evolution period followed by 13C-TOCSY mixing for aliphatic 1H resonance assignment without reliance on 1H detection.