Communication
Bicelle crystallization: a new method for crystallizing membrane proteins yields a monomeric bacteriorhodopsin structure1

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Abstract

Obtaining crystals of membrane proteins that diffract to high resolution remains a major stumbling block in structure determination. Here we present a new method for crystallizing membrane proteins from a bicelle forming lipid/detergent mixture. The method is flexible and simple to use. As a test case, bacteriorhodopsin (bR) from Halobacterium salinarum was crystallized from a bicellar solution, yielding a new bR crystal form. The crystals belong to space group P21 with unit cell dimensions of a = 45.0 Å, b = 108.9 Å, c = 55.9 Å, β = 113.58° and a dimeric asymmetric unit. The structure was solved by molecular replacement and refined at 2.0 Å resolution. In all previous bR structures the protein is organized as a parallel trimer, but in the crystals grown from bicelles, the individual bR subunits are arranged in an antiparallel fashion.

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Acknowledgments

The authors thank Todd Yeates for helpful discussions concerning the possibility of twinning, Sarah Yohannan, Duilio Cascio, Mike Sawaya and the staff at BNL beamline X8C for help with data collection. We also thank Aaron Chamberlain, Hoang Tran, Ranjini Ramachander, Sarah Yohannan, Marisa Baron and Megan Mathews for comments on the manuscript. This work was supported by NIH grants RO1 GM59164 and RO1 GM63919. J.U.B. is a Leukemia and Lymphoma Society Scholar.

References (30)

  • J Deisenhofer et al.

    Structure of the protein subunits in the photosynthetic reaction center of Rhodopseudomonas viridis at 3 Ångstrom resolution

    Nature

    (1985)
  • H Michel

    General and practical aspects of membrane protein crystallization

  • M Roth et al.

    Detergent structure in crystals of a bacterial photosynthetic reaction center

    Nature

    (1989)
  • J Deisenhofer et al.

    The photosynthetic reaction center from the purple bacterium Rhodopseudomonas viridis

    Science

    (1989)
  • S Iwata et al.

    Structure at 2.8 Å resolution of cytochrome c oxidase from Paracoccus denitrificans

    Nature

    (1995)
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    Edited by D. Rees

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    Present address: James U. Bowie, Boyer Hall, UCLA, 611 Charles E. Young Dr. East, Los Angeles, CA 90095-1570, USA.

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