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Structural alterations for proton translocation in the M state of wild-type bacteriorhodopsin

Nature volume 406, pages 649–653 (2000)Cite this article

Abstract

The transport of protons across membranes is an important process in cellular bioenergetics. The light-driven proton pump bacteriorhodopsin is the best-characterized protein providing this function. Photon energy is absorbed by the chromophore retinal, covalently bound to Lys 216 via a protonated Schiff base. The light-induced all-trans to 13-cis isomerization of the retinal results in deprotonation of the Schiff base followed by alterations in protonatable groups within bacteriorhodopsin. The changed force field induces changes, even in the tertiary structure1,2,3, which are necessary for proton pumping. The recent report4 of a high-resolution X-ray crystal structure for the late M intermediate of a mutant bacteriorhopsin (with Asp 96→Asn) displays the structure of a proton pathway highly disturbed by the mutation. To observe an unperturbed proton pathway, we determined the structure of the late M intermediate of wild-type bacteriorhodopsin (2.25 Å resolution). The cytoplasmic side of our M2 structure shows a water net that allows proton transfer from the proton donor group Asp 96 towards the Schiff base. An enlarged cavity system above Asp 96 is observed, which facilitates the de- and reprotonation of this group by fluctuating water molecules in the last part of the cycle.

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Figure 1: Image of the proton pathway of the BR model (purple) and the M2-state model (yellow).
Figure 2: Details of the structural differences between the ground state and the M2 intermediate.
Figure 3: Stereo images of the M2-state model of wild-type bacteriorhodopsin.
Figure 4: Comparison of the late M-state models of wild-type bacteriorhodopsin (yellow) with the mutant Asp96→Asn (green; ref.

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Acknowledgements

We thank E. Landau, J. Rosenbusch, J. Heberle, V. Gordeliy, J. Granzin and J. Labahn for support and discussions, and A. Cousin for preparation of purple membranes. This work was supported by EU-BIOTECH and the Deutsche Forschungsgemeinschaft, Sfb 189 (H.J.S. and G.B.), the US Department of Energy, Office of Health and Enviromental Research (J.B.), and OTKA (P.O.).

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Authors and Affiliations

  1. Institute of Structural Biology, Research Centre Jülich, Jülich, 52425, Germany

    Hans Jürgen Sass, Georg Büldt, Ralf Gessenich, Dominic Hehn, Dirk Neff & Ramona Schlesinger

  2. Biophysics Group, Mail Stop D454, Los Alamos Laboratory, Los Alamos, 87545, New Mexico, USA

    Joel Berendzen

  3. Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, PO Box 521, Szeged, 6701, Hungary

    Pal Ormos

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  1. Hans Jürgen Sass
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Correspondence to Georg Büldt.

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Sass, H., Büldt, G., Gessenich, R. et al. Structural alterations for proton translocation in the M state of wild-type bacteriorhodopsin. Nature 406, 649–653 (2000). https://doi.org/10.1038/35020607

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