High resolution 13C-detected solid-state NMR spectroscopy of a deuterated protein
Abstract 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, 13Cā?², 13CĪ± and 13CĪ² sites are resolved in 13Cā??13C and 15Nā??13C 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 13CĪ±
T 2ā?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 proteinā??protein interactions.
- Content Type Journal Article
- DOI 10.1007/s10858-010-9442-8
- Authors
- Ming Tang, Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA
- Gemma Comellas, Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA
- Leonard J. Mueller, Department of Chemistry, University of California, Riverside, CA 92521, USA
- Chad M. Rienstra, Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA
Source: Journal of Biomolecular NMR