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Two-Dimensional Crystalline Array Formation of Glucuronide Transporter from Escherichia coli by the Use of Polystyrene Beads for Detergent Removal

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Abstract

n-Dodecyl-β-d-maltoside solubilized glucuronide transporter (GusB), the product of gusB gene from Escherichia coli, was treated with Bio-Beads as an agent for removing the detergent from a micellar solution under suitable combination with dimyristoylphosphatidylcholine. Optimizing conditions led to a two-dimensional crystalline array formation of GusB. The crystalline arrays appear to have a hexagonal lattice with layer group P6, the unit cell dimensions of a = b = 13.8 nm and γ = 120°. Each stain-protruding periodic unit showed approximately 11.8 ± 0.3 nm in a diameter in the inverse Fourier-filtered image to have formed with pentameric GusB (5 × 49.7 kDa).

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Acknowledgments

The author would like to express thanks to Prof. R. M. Glaeser for stimulating discussion, and Drs. F. M. Hendrickson and K. S. Kim for discussions and critical reading. The part of the research was financially supported by the Overseas Research Fellowship from Japan Science and Technology Corporation.

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Authors and Affiliations

  1. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central-6 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Noriyuki Ishii

  2. Life Science Division, Lawrence Berkeley National Laboratory, Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720, USA

    Noriyuki Ishii

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  1. Noriyuki Ishii
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Correspondence to Noriyuki Ishii.

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Ishii, N. Two-Dimensional Crystalline Array Formation of Glucuronide Transporter from Escherichia coli by the Use of Polystyrene Beads for Detergent Removal. J Membrane Biol 246, 199–207 (2013). https://doi.org/10.1007/s00232-012-9521-8

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  • DOI: https://doi.org/10.1007/s00232-012-9521-8

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