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

High resolution 13C-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 15N, 13C', 13Cα and 13Cβ sites are resolved in 13C-13C and 15N-13C spectra, with significant improvement in T2 relaxation times and resolution at high magnetic field (750 MHz). The comparison of echo T2 values between deuterated and protonated GB1 at various spinning rates and under different decoupling schemes indicates that 13Cα T2' 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.