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

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, 13Câ?², 13Cα and 13Cβ sites are resolved in 13Câ??13C and 15Nâ??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 13Cα T 2â?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 proteinâ??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

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR