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The neuronal t-SNARE complex is a parallel four-helix bundle

Nature Structural Biology volume 8, pages 308–311 (2001)Cite this article

Abstract

Assembly of the soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) complex is an essential step for neurotransmitter release in synapses. The presynaptic plasma membrane associated proteins (t-SNAREs), SNAP-25 (synaptosome-associated protein of 25,000 Da) and syntaxin 1A may form an intermediate complex that later binds to vesicle-associated membrane protein 2 (VAMP2). Using spin labeling electron paramagnetic resonance (EPR), we found that the two t-SNARE proteins assemble into a parallel four-helix bundle that consists of two identical syntaxin 1A components and the N-terminal and C-terminal domains of SNAP-25. Although the structure is generally similar to that of the final SNARE complex, the middle region of the helical bundle appears more flexible in the t-SNARE complex. Such flexibility might facilitate interactions between VAMP2 and the t-SNARE complex.

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Figure 1: Spin labeling EPR analysis of SNARE complexes.
Figure 2: Schematic representation of SNARE complexes showing interspin distances.
Figure 3: Selected low temperature (−140 °C) integrated-mode EPR spectra of SNARE complexes prepared with doubly labeled SNAP-25[N].
Figure 4: Flexibility in the central region of the t-SNARE complex revealed by EPR lineshape analysis.

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Acknowledgements

The authors thank J. Aslan and C. Chan for technical assistance, D. King for mass spectrometry analysis and L.C. Gonzalez, Jr. for helpful discussions and critical reading of the manuscript. Support for this research was provided by the National Institutes of Health (M.K.B., Y.-K.S.), the McKnight Fund for Neuroscience (M.K.B.) and a Searle Scholarship (Y.-K.S.)

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Author notes

  1. Michelle A. Poirier

    Present address: Department of Psychiatry, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA

  2. Wenzhong Xiao and Michelle A. Poirier: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of California, Berkeley, 94720, California, USA

    Wenzhong Xiao

  2. Department of Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Michelle A. Poirier & Mark K. Bennett

  3. Department of Biochemistry and Biophysics, Iowa State Univeristy, Ames, 50011, Iowa, USA

    Yeon-Kyun Shin

  4. Rigel Pharmaceuticals, Inc., 240 E. Grand Ave., South San Francisco, 94080, California, USA

    Mark K. Bennett

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  1. Wenzhong Xiao
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  4. Yeon-Kyun Shin
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Correspondence to Yeon-Kyun Shin.

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Xiao, W., Poirier, M., Bennett, M. et al. The neuronal t-SNARE complex is a parallel four-helix bundle. Nat Struct Mol Biol 8, 308–311 (2001). https://doi.org/10.1038/86174

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