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Hemifusion in SNARE-mediated membrane fusion

Nature Structural & Molecular Biology volume 12, pages 417–422 (2005)Cite this article

Abstract

SNAREs are essential for intracellular membrane fusion. Using EPR, we determined the structure of the transmembrane domain (TMD) of the vesicle (v)-SNARE Snc2p involved in trafficking in yeast. Structural features of the TMD were used to design a v-SNARE mutant in which about half of the TMD was deleted. Liposomes containing this mutant induced outer leaflet mixing but not inner leaflet mixing when incubated with liposomes containing target membrane (t)-SNAREs. Hemifusion was also detected with wild-type SNAREs when low protein concentrations were reconstituted. Thus, these results show that SNARE-mediated fusion can transit through a hemifusion intermediate.

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Figure 1: Site-directed spin labeling and the EPR analysis of Snc2p.
Figure 2: Structural determination of the Snc2p TMD using EPR.
Figure 3: Total and inner leaflet lipid mixing for Snc2p and its mutants.
Figure 4: Total and inner leaflet lipid mixing assays at low protein concentrations.

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Acknowledgements

Support for this work was provided by the US National Institutes of Health (Y-.K.S) and the US National Science Foundation and the Robert A. Welch Foundation (J.A.M.).

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Author notes

  1. Yibin Xu and Fan Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, 50011, Iowa, USA

    Yibin Xu, Fan Zhang, Zengliu Su & Yeon-Kyun Shin

  2. Department of Biochemistry and Cell Biology, Rice University, Houston, 77251-1892, Texas, USA

    James A McNew

Authors
  1. Yibin Xu
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  2. Fan Zhang
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  4. James A McNew
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Correspondence to Yeon-Kyun Shin.

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Supplementary information

Supplementary Fig. 1

Lipid-mixing assay for SNARE-free PE-containing vesicles (PDF 287 kb)

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Xu, Y., Zhang, F., Su, Z. et al. Hemifusion in SNARE-mediated membrane fusion. Nat Struct Mol Biol 12, 417–422 (2005). https://doi.org/10.1038/nsmb921

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