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Vam3p structure reveals conserved and divergent properties of syntaxins

Nature Structural Biology volume 8, pages 258–264 (2001)Cite this article

Abstract

Syntaxins and Sec1/munc18 proteins are central to intracellular membrane fusion. All syntaxins comprise a variable N-terminal region, a conserved SNARE motif that is critical for SNARE complex formation, and a transmembrane region. The N-terminal region of neuronal syntaxin 1A contains a three-helix domain that folds back onto the SNARE motif forming a 'closed' conformation; this conformation is required for munc18-1 binding. We have examined the generality of the structural properties of syntaxins by NMR analysis of Vam3p, a yeast syntaxin essential for vacuolar fusion. Surprisingly, Vam3p also has an N-terminal three-helical domain despite lacking apparent sequence homology with syntaxin 1A in this region. However, Vam3p does not form a closed conformation and its N-terminal domain is not required for binding to the Sec1/munc18 protein Vps33p, suggesting that critical distinctions exist in the mechanisms used by syntaxins to govern different types of membrane fusion.

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Figure 1: Vam3p contains an autonomously folded N-terminal domain.
Figure 2: Structure of the Vam3N domain.
Figure 3: Ribbon diagrams showing superpositions of the Vam3N domain (blue) and the N-terminal domain of syntaxin 1A (orange).
Figure 4: Comparison of the sequence and domain structure of Vam3p with those of plasma membrane syntaxins.
Figure 5: 1H-15N HSQC spectra of Vam3p fragments.
Figure 6: The Vam3p N-terminal domain is not required for HA-Vps33p binding.

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Acknowledgements

We thank Y. Gao for technical assistance, B. Horazdovsky for reagents and fruitful discussions, and L. Kay for providing pulse sequences. This work was supported by an Established Investigator grant from the American Heart Association and by an NIH grant to JR.

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Authors and Affiliations

  1. Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, 75390, Texas, USA

    Irina Dulubova & Josep Rizo

  2. Department of Molecular Genetics and Howard Hughes Medical Institute, Center for Basic Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, 75390, Texas, USA

    Tomohiro Yamaguchi, Yun Wang & Thomas C. Südhof

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Correspondence to Josep Rizo.

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Dulubova, I., Yamaguchi, T., Wang, Y. et al. Vam3p structure reveals conserved and divergent properties of syntaxins. Nat Struct Mol Biol 8, 258–264 (2001). https://doi.org/10.1038/85012

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