title: Structures of yeast vesicle trafficking proteins.
abstract: In protein transport between organelles, interactions of v- and t-SNARE proteins are required for fusion of protein-containing vesicles with appropriate target compartments. Mammalian SNARE proteins have been observed to interact with NSF and SNAP, and yeast SNAREs with yeast homologues of NSF and SNAP proteins. This observation led to the hypothesis that, despite low sequence homology, SNARE proteins are structurally similar among eukaryotes. SNARE proteins can be classified into two groups depending on whether they interact with SNARE binding partners via conserved glutamine (Q-SNAREs) or arginine (R-SNAREs). Much of the published structural data available is for SNAREs involved in exocytosis (either in yeast or synaptic vesicles). This paper describes circular dichroism, Fourier transform infrared spectroscopy, and dynamic light scattering data for a set of yeast v- and t-SNARE proteins, Vti1p and Pep12p, that are Q-SNAREs involved in intracellular trafficking. Our results suggest that the secondary structure of Vti1p is highly alpha-helical and that Vti1p forms multimers under a variety of solution conditions. In these respects, Vti1p appears to be distinct from R-SNARE proteins characterized previously. The alpha-helicity of Vti1p is similar to that of Q-SNARE proteins characterized previously. Pep12p, a Q-SNARE, is highly alpha-helical. It is distinct from other Q-SNAREs in that it forms dimers under many of the solution conditions tested in our experiments. The results presented in this paper are among the first to suggest heterogeneity in the functioning of SNARE complexes.
authors.affiliation: Department of Biochemistry & Molecular Biology, University of Georgia, Athens 30602, USA.
authors.forename: T
authors.initials: T
authors.lastname: Tishgarten
authors.forename: F F
authors.initials: FF
authors.lastname: Yin
authors.forename: K M
authors.initials: KM
authors.lastname: Faucher
authors.forename: R A
authors.initials: RA
authors.lastname: Dluhy
authors.forename: T R
authors.initials: TR
authors.lastname: Grant
authors.forename: G
authors.initials: G
authors.lastname: Fischer von Mollard
authors.forename: T H
authors.initials: TH
authors.lastname: Stevens
authors.forename: L A
authors.initials: LA
authors.lastname: Lipscomb
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry*; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry; D008027:Light; D008322:Mammals; D008565:Membrane Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D012542:Scattering, Radiation; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; C081224:PEP12 protein, S cerevisiae; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C106736:VTI1 protein, S cerevisiae; D033921:Vesicular Transport Proteins