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Cocrystal structure of synaptobrevin-II bound to botulinum neurotoxin type B at 2.0 Å resolution

Nature Structural Biology volume 7, pages 687–692 (2000)Cite this article

A Retraction to this article was published on 01 July 2009

Abstract

Botulinum neurotoxin serotype B is a zinc protease that disrupts neurotransmitter release by cleaving synaptobrevin-II (Sb2), one of three SNARE proteins involved in neuronal synaptic vesicle fusion. The three-dimensional crystal structure of the apo botulinum neurotoxin serotype B catalytic domain (BoNT/B-LC) has been determined to 2.2 Å resolution, and the complex of cleaved Sb2 with the catalytic domain (Sb2–BoNT/B-LC) has been determined to 2.0 Å resolution. A comparison of the holotoxin catalytic domain and the isolated BoNT/B-LC structure shows a rearrangement of three active site loops. This rearrangement exposes the BoNT/B active site. The Sb2–BoNT/B-LC structure illustrates two distinct binding regions, which explains the specificity of each botulinum neurotoxin for its synaptic vesicle protein. This observation provides an explanation for the proposed cooperativity between binding of full-length substrate and catalysis and suggest a mechanism of synaptobrevin proteolysis employed by the clostridial neurotoxins.

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Figure 1: Structures of BoNT/A holotoxin and BoNT/B-LC.
Figure 2: Stereo view showing the electron density contoured at 1.2σ for the cleaved Sb2 peptides (purple).
Figure 3: Molecular and schematic representations of the interactions formed between the bound Sb2 products and BoNT/B-LC.
Figure 4: The proposed reaction mechanism for synaptobrevin proteolysis by botulinum neurotoxin.
Figure 5: The SNARE complex showing Sb2 residues important for toxin binding.

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Acknowledgements

We dedicate this paper to H. Niemann, who provided the original light chain constructs to us, and who recently passed away. We thank P. Kuhn and the SSRL staff for their assistance at beamlines 9-1 and 9-2 at the Stanford Synchrotron Radiation Laboratory and we thank M. Adler and J. Nicholson for the synaptobrevin peptide used in the co-crystal structure. We also greatly appreciate the neurotoxin provided by B. DasGupta, B. Tepp, E. Johnson, and M. Goodenough used in previous structural studies and their helpful discussions. Financial support for this research was provided by the U.S. Army and the NN20 program of the U.S. Department of Energy.

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  1. Department of Chemistry and Molecular Biology, The Scripps Research Institute, La Jolla, 92037, California, USA

    Michael A. Hanson & Raymond C. Stevens

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  1. Michael A. Hanson
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  2. Raymond C. Stevens
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Correspondence to Raymond C. Stevens.

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Hanson, M., Stevens, R. Cocrystal structure of synaptobrevin-II bound to botulinum neurotoxin type B at 2.0 Å resolution. Nat Struct Mol Biol 7, 687–692 (2000). https://doi.org/10.1038/77997

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