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Advances towards resonance assignments for uniformly—13C, 15N enriched bacteriorhodopsin at 18.8 T in purple membranes

Journal of Biomolecular NMR volume 41, pages 1–4 (2008)Cite this article

Abstract

Solid state NMR spectra from uniformly 13C, 15N enriched bacteriorhodospin (bR) purified from H. salinarium were acquired at 18.8 T using magic angle spinning methods. Isolated resonances of 2D 13C−13C spectra exhibited 0.50–0.55 ppm line-widths. Several amino acid types could be assigned, and at least 12 out of 15 Ile peaks could be resolved clearly and identified based on their characteristic chemical shifts and connectivities. This study confirms that high resolution solid state NMR spectra can be obtained for a 248 amino acid uniformly labeled membrane protein in its native membrane environment and indicates that site-specific assignments are likely to be feasible with heteronuclear multidimensional spectra.

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Fig. 1
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Abbreviations

NMR:

Nuclear magnetic resonance

bR:

Bacteriorhodospin

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Acknowledgements

We acknowledge financial support from EPSRC (under grants GR/R99393/01 and EP/C015452/1), Bionanotechnology IRC, Magnex Scientific and Varian Inc., and the advice from the staff of the ILL-EMBL Deuteration Laboratory. This work has benefited from the activities of the DLAB & JRA7 consortia funded by the EU under contracts HPRI–2001–50065 and RII3–CT–2003–505925.

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Authors and Affiliations

  1. Biomembrane Structure Unit, Department of Biochemistry, Oxford University, South Parks Road, Oxford, OX1 3QU, UK

    Krisztina Varga, Lubica Aslimovska & Anthony Watts

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  1. Krisztina Varga
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  2. Lubica Aslimovska
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  3. Anthony Watts
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Correspondence to Anthony Watts.

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Varga, K., Aslimovska, L. & Watts, A. Advances towards resonance assignments for uniformly—13C, 15N enriched bacteriorhodopsin at 18.8 T in purple membranes. J Biomol NMR 41, 1–4 (2008). https://doi.org/10.1007/s10858-008-9235-5

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  • DOI: https://doi.org/10.1007/s10858-008-9235-5

Keywords

  • Solid state NMR
  • Bacteriorhodospin
  • Assignment
  • Membrane protein