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SNARE assembly and disassembly exhibit a pronounced hysteresis

Nature Structural Biology volume 9, pages 144–151 (2002)Cite this article

Abstract

SNARE proteins are essential for intracellular membrane fusion of eukaryotes. Their assembly into stable four-helix bundles bridges membranes and may provide the energy for initiating membrane fusion. In vitro, assembly of soluble SNARE fragments is accompanied by major structural rearrangements that can be described as a folding reaction. The pathways and the thermodynamics of SNARE protein interactions, however, are not known. Here we report that assembly and dissociation of two distantly related SNARE complexes exhibit a marked hysteresis. The assembled and disassembled native states are separated by a kinetic barrier and cannot equilibrate on biologically relevant timescales. We suggest that the hysteresis is a hallmark of all SNARE complexes and that complex assembly and disassembly follow different pathways that may be independently controlled.

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Figure 1: Unfolding and refolding transitions of the synaptic SNARE complex exhibited a marked hysteresis.
Figure 2: Hysteresis in the unfolding and folding transition of the synaptic SNARE complex visualized by SDS-PAGE.
Figure 3: Unfolding and refolding transitions of an endosomal SNARE complex are noncoincident.
Figure 4: Equilibrium unfolding of the syntaxin–SNAP-25 complex monitored by CD spectroscopy.
Figure 5: Unfolding kinetics of the synaptic complex monitored by CD spectroscopy at 220 nm.
Figure 6: Biphasic refolding kinetics of the synaptic SNARE complex suggest assembly via a syntaxin–SNAP-25 intermediate.

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Acknowledgements

The authors are very much indebted to K. Fiebig for pointing out in the beginning of the study that the observed hysteresis could be due to a folding intermediate. We thank M. Margittai, R. Langen, D. Bruns, T. Heimburg, S. Pabst, V. Knecht, G. Schröder and H. Grubmüller for stimulating discussions, suggestions and critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, AM Fassberg 11, Göttingen, 37077, Germany

    Dirk Fasshauer, Wolfram Antonin & Reinhard Jahn

  2. Department of Molecular Biology, Max-Planck-Institute for Biophysical Chemistry, AM Fassberg 11, Göttingen, 37077, Germany

    Vinod Subramaniam

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Correspondence to Dirk Fasshauer.

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Fasshauer, D., Antonin, W., Subramaniam, V. et al. SNARE assembly and disassembly exhibit a pronounced hysteresis. Nat Struct Mol Biol 9, 144–151 (2002). https://doi.org/10.1038/nsb750

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