Journal of Biomolecular NMR

, Volume 48, Issue 2, pp 103–111

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

  • Ming Tang
  • Gemma Comellas
  • Leonard J. Mueller
  • Chad M. Rienstra
Article

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, 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 13T2′ 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.

Keywords

Deuterated protein Solid-state NMR 13C-detected spectra Deuterium effect T2′ relaxation rates 

Abbreviations used

DARR

Dipolar assisted rotational resonance

GB1

The beta-1 immunoglobulin binding domain of protein G

MAS

Magic-angle spinning

SSNMR

Solid-state nuclear magnetic resonance

CTUC COSY

Constant time uniform-sign cross-peak correlation spectroscopy

IPAP

In-phase anti-phase

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Ming Tang
    • 1
  • Gemma Comellas
    • 2
  • Leonard J. Mueller
    • 4
  • Chad M. Rienstra
    • 1
    • 2
    • 3
  1. 1.Department of ChemistryUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Center for Biophysics and Computational BiologyUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  3. 3.Department of BiochemistryUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  4. 4.Department of ChemistryUniversity of CaliforniaRiversideUSA

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