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Crystal structure of Staphylococcal LukF delineates conformational changes accompanying formation of a transmembrane channel

Nature Structural Biology volume 6, pages 134–140 (1999)Cite this article

Abstract

Staphylococcal LukF, LukS, HγII, and α–hemolysin are self–assembling, channel–forming proteins related in sequence and function. In the α–hemolysin heptamer, the channel–forming β–strands and the amino latch make long excursions from the protomer core. Here we report the crystal structure of the water soluble form of LukF. In the LukF structure the channel–forming region folds into an amphipathic, three–strand β–sheet and the amino latch forms a β–strand extending a central β–sheet. The LukF structure illustrates how a channel–forming toxin masks protein–protein and protein–membrane interfaces prior to cell binding and assembly, and together with the α–hemolysin heptamer structure, they define the end points on the pathway of toxin assembly.

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Figure 1: LukF structure.
Figure 2: a, Ribbon representation of the LukF monomer numbered and colored according to the homologous strands and helices from the αHL protomer (ref. 10, Fig 3a).
Figure 3: Superpositions of LukF and a single protomer taken from the αHL heptamer structure.
Figure 4: Mechanism of assembly for αHL using the structure of LukF as a model for the water soluble and membrane–bound monomers, as well as for the pre–pore subunits.

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Acknowledgements

The assistance of G.–Q. Chen, Y. Sun and N. Armstrong in synchrotron data collection and the help from S. Galdiero in data collection and processing are greatly appreciated. Superb support of the X–ray laboratory at Columbia by J. Lidestri is acknowledged as is general assistance from members of the Hendrickson group. Suggestions from L. Shapiro and D. Leahy on the growth of B834 E. coli and advice from T. Terwilliger on using Solve are also acknowledged. The MAD data collection component of this research was conducted at the Cornell High Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation under award DMR–9311772, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by an award from the NIH. The diffraction data from the LukF–DiC3PC crystals was measured at 14–BM–D (BioCARS) at the Advanced Photon Source and we thank the beamline staff for their assistance. This work was also supported by the NIH (E.G.) and a Grant–In–Aid for Scientific Research from the Ministry of Education, Science Sport and Culture of Japan to YK. E.G. is a NSF Young Investigator and the recipient of an Alfred P. Sloan Research Fellowship.

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  1. Department of Biochemistry and Molecular Biophysics, Columbia University, 650 W. 168th Street, New York, 10032, New York, USA

    Rich Olson & Eric Gouaux

  2. Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoko University, 1-1 Amamiya-machi, Tsutsumi-dori, Sendai, 981, Aoba-ku, Japan

    Hirofumi Nariya, Kenji Yokota & Yoshiyuki Kamio

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Correspondence to Eric Gouaux.

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Olson, R., Nariya, H., Yokota, K. et al. Crystal structure of Staphylococcal LukF delineates conformational changes accompanying formation of a transmembrane channel. Nat Struct Mol Biol 6, 134–140 (1999). https://doi.org/10.1038/5821

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