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Characterisation of Schiff base and chromophore in green proteorhodopsin by solid-state NMR

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Abstract

The proteorhodopsin family consists of hundreds of homologous retinal containing membrane proteins found in bacteria in the photic zone of the oceans. They are colour tuned to their environment and act as light-driven proton pumps with a potential energetic and regulatory function. Precise structural details are still unknown. Here, the green proteorhodopsin variant has been selected for a chemical shift analysis of retinal and Schiff base by solid-state NMR. Our data show that the chromophore exists in mainly all-trans configuration in the proteorhodopsin ground state. The optical absorption maximum together with retinal and Schiff base chemical shifts indicate a strong interaction network between chromophore and opsin.

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Abbreviations

bR:

Bacteriorhodopsin

PR:

Proteorhodopsin

wt:

Wild type

PSB:

Protonated Schiff base

DA:

Dark adapted

LA:

Light adapted

MAS:

Magic angle sample spinning

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Acknowledgments

This work has been financially supported by SFB 472 (“Molecular Bioenergetics”). Cells and plasmids were kindly provided by E. Bamberg, Frankfurt and M. Engelhard, Dortmund. The 10,11-13C2 retinal was a generous gift of Marina Carravetta and Malcolm Levitt, Southampton. Ute Hellmich and Christoph Kaiser are acknowledged for helpful discussions.

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Author notes

  1. Mark Lorch

    Present address: Department of Chemistry, University of Hull, Hull, UK

Authors and Affiliations

  1. Institute for Biophysical Chemistry, Centre for Biomolecular Magnetic Resonance, J. W. Goethe University, Max von Laue Str. 9, 60438, Frankfurt am Main, Germany

    Nicole Pfleger, Mark Lorch, Andreas C. Woerner, Sarika Shastri & Clemens Glaubitz

Authors
  1. Nicole Pfleger
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  2. Mark Lorch
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  3. Andreas C. Woerner
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  4. Sarika Shastri
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Corresponding author

Correspondence to Clemens Glaubitz.

Additional information

Mark Lorch and Andreas C. Woerner contributed equally to this work.

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Pfleger, N., Lorch, M., Woerner, A.C. et al. Characterisation of Schiff base and chromophore in green proteorhodopsin by solid-state NMR. J Biomol NMR 40, 15–21 (2008). https://doi.org/10.1007/s10858-007-9203-5

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  • DOI: https://doi.org/10.1007/s10858-007-9203-5

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