Furgason, M.L.M., MacDonald, C., Shanks, S.G., Ryder, S.P., Bryant, N.J. and Munson, M. (2009) The N-terminal peptide of the syntaxin Tlg2p modulates binding of its closed conformation to Vps45p. Proceedings of the National Academy of Sciences of the United States of America, 106(34), pp. 14303-14308. (doi: 10.1073/pnas.0902976106) (PMID:19667197) (PMCID:PMC2732825)
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Abstract
The Sec1/Munc18 (SM) protein family regulates intracellular trafficking through interactions with individual SNARE proteins and assembled SNARE complexes. Revealing a common mechanism of this regulation has been challenging, largely because of the multiple modes of interaction observed between SM proteins and their cognate syntaxin-type SNAREs. These modes include binding of the SM to a closed conformation of syntaxin, binding to the N-terminal peptide of syntaxin, binding to assembled SNARE complexes, and/or binding to nonsyntaxin SNAREs. The SM protein Vps45p, which regulates endosomal trafficking in yeast, binds the conserved N-terminal peptide of the syntaxin Tlg2p. We used size exclusion chromatography and a quantitative fluorescent gel mobility shift assay to reveal an additional binding site that does not require the Tlg2p N-peptide. Characterization of Tlg2p mutants and truncations indicate that this binding site corresponds to a closed conformation of Tlg2p. Furthermore, the Tlg2p N-peptide competes with the closed conformation for binding, suggesting a fundamental regulatory mechanism for SM-syntaxin interactions in SNARE assembly and membrane fusion.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Bryant, Dr Nia and Shanks, Dr Scott |
| Authors: | Furgason, M.L.M., MacDonald, C., Shanks, S.G., Ryder, S.P., Bryant, N.J., and Munson, M. |
| Subjects: | Q Science > QH Natural history > QH345 Biochemistry |
| College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Journal Name: | Proceedings of the National Academy of Sciences of the United States of America |
| ISSN: | 0027-8424 |
| ISSN (Online): | 1091-6490 |
| Published Online: | 25 August 2009 |
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