A new NMR experiment is described for recording NOEs from Val and Ile methyl groups in 15N, 13C-labeled or methyl-protonated, 15N, 13C, 2H-labeled proteins that offers far superior resolution than conventional 3D 13C-edited NOESY data sets. Resolution is achieved by recording both the C(β) and C(γ) (Val) or C(γ2) (Ile) chemical shifts as well as the chemical shift of the destination proton, and a strategy is introduced for refocusing homonuclear carbon couplings during the constant-time evolution of C(β) carbon magnetization. The utility of the method is demonstrated with applications on a 160- residue fully proton areal 15N, 13C-labeled, dNumb PTB domain- peptide complex and a methyl protonated, highly deuterated 15N, 13C-labeled complex of maltose binding protein and β-cyclodextrin (42 kDa).
ASJC Scopus subject areas
- Colloid and Surface Chemistry