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Structure of α-latrotoxin oligomers reveals that divalent cation-dependent tetramers form membrane pores

Nature Structural Biology volume 7, pages 48–53 (2000)Cite this article

Abstract

We report here the first three-dimensional structure of α-latrotoxin, a black widow spider neurotoxin, which forms membrane pores and stimulates secretion in the presence of divalent cations. We discovered that α-latrotoxin exists in two oligomeric forms: it is dimeric in EDTA but forms tetramers in the presence of Ca2+ or Mg2+. The dimer and tetramer structures were determined independently at 18 Å and 14 Å resolution, respectively, using cryo-electron microscopy and angular reconstitution. The α-latrotoxin monomer consists of three domains. The N- and C-terminal domains have been identified using antibodies and atomic fitting. The C4-symmetric tetramers represent the active form of α-latrotoxin; they have an axial channel and can insert into lipid bilayers with their hydrophobic base, providing the first model of α-latrotoxin pore formation.

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Figure 1: Formation and properties of LTX dimers and tetramers.
Figure 2: Cryoelectron microscopy of LTX.
Figure 3: The 3D reconstruction of the LTX oligomers.
Figure 4: Analysis of the LTX monomer structure.
Figure 5: Cryo-EM of membrane-incorporated LTX.

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Acknowledgements

We thank R. Weinzierl for the use of the DLS spectrometer. The work was funded by a Wellcome Trust Senior European Research Fellowship (to Y.A.U.) and BBSRC and EC grants (to M.v.H.).

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Authors and Affiliations

  1. Biochemistry Department, Imperial College, Exhibition Road, London, SW7 2AY, UK

    Elena V. Orlova, M. Atiqur Rahman, Brent Gowen, Kirill E. Volynski, Anthony C. Ashton, Catherine Manser, Marin van Heel & Yuri A. Ushkaryov

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Correspondence to Yuri A. Ushkaryov.

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Orlova, E., Rahman, M., Gowen, B. et al. Structure of α-latrotoxin oligomers reveals that divalent cation-dependent tetramers form membrane pores. Nat Struct Mol Biol 7, 48–53 (2000). https://doi.org/10.1038/71247

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