High resolution 13C-detected solid-state NMR spectroscopy of a deuterated protein

High resolution C-13-detected solid-state NMR spectra of the deuterated beta-1 immunoglobulin binding domain of the protein G (GB1) have been collected to show that all N-15, C-13', C-13 alpha and C-13 beta sites are resolved in C-13-C-13 and N-15-C-13 spectra, with significant improvement in T (2) relaxation times and resolution at high magnetic field (750 MHz). The comparison of echo T (2) values between deuterated and protonated GB1 at various spinning rates and under different decoupling schemes indicates that C-13 alpha T (2)' times increase by almost a factor of two upon deuteration at all spinning rates and under moderate decoupling strength, and thus the deuteration enables application of scalar-based correlation experiments that are challenging from the standpoint of transverse relaxation, with moderate proton decoupling. Additionally, deuteration in large proteins is a useful strategy to selectively detect polar residues that are often important for protein function and protein-protein interactions.