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Journal of Biomolecular NMR

, 45:319 | Cite as

Characterization of different water pools in solid-state NMR protein samples

  • Anja BöckmannEmail author
  • Carole Gardiennet
  • René Verel
  • Andreas Hunkeler
  • Antoine Loquet
  • Guido Pintacuda
  • Lyndon Emsley
  • Beat H. Meier
  • Anne LesageEmail author
Article

Abstract

We observed and characterized two distinct signals originating from different pools of water protons in solid-state NMR protein samples, namely from crystal water which exchanges polarization with the protein (on the NMR timescale) and is located in the protein-rich fraction at the periphery of the magic-angle spinning (MAS) sample container, and supernatant water located close to the axis of the sample container. The polarization transfer between the water and the protein can be probed by two-dimensional exchange spectroscopy, and we show that the supernatant water does not interact with protein on the timescale of the experiments. The two water pools have different spectroscopic properties, including resonance frequency, longitudinal, transverse and rotating frame relaxation times. The supernatant water can be removed almost completely physically or can be frozen selectively. Both measures lead to an enhancement of the quality factor of the probe circuit, accompanied by an improvement of the experimental signal/noise, and greatly simplify solvent-suppression by substantially reducing the water signal. We also present a tool, which allows filling solid-state NMR sample containers in a more efficient manner, greatly reducing the amount of supernatant water and maximizing signal/noise.

Keywords

Solid-state NMR Water–protein polarization transfer Microcrystalline protein Chemical exchange 

Notes

Acknowledgement

We thank Michel Juy for the picture of the Crh proteins arranged in the unit cell. This work was funded in part by CNRS, the French Research Ministry (ANR JCJC JC05_44957, ANR PCV 07 PROTEIN MOTION), the Swiss National Science Foundation (SNF) and the ETH Zurich.

Supplementary material

10858_2009_9374_MOESM1_ESM.pdf (658 kb)
(PDF 659 kb)

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Anja Böckmann
    • 1
    Email author
  • Carole Gardiennet
    • 1
  • René Verel
    • 2
  • Andreas Hunkeler
    • 2
  • Antoine Loquet
    • 1
  • Guido Pintacuda
    • 3
  • Lyndon Emsley
    • 3
  • Beat H. Meier
    • 2
  • Anne Lesage
    • 3
    Email author
  1. 1.Institut de Biologie et Chimie des ProtéinesUniversité de Lyon, UMR 5086 CNRS/UCB-Lyon 1LyonFrance
  2. 2.Physical ChemistryETH ZürichZurichSwitzerland
  3. 3.Université de Lyon, CNRS/ENS Lyon/UCB-Lyon 1VilleurbanneFrance

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