title: Epsin N-terminal homology domains bind on opposite sides of two SNAREs.
abstract: SNARE proteins are crucial for membrane fusion in vesicular transport. To ensure efficient and accurate fusion, SNAREs need to be sorted into different budding vesicles. This process is usually regulated by specific recognition between SNAREs and their adaptor proteins. How different pairs of SNAREs and adaptors achieve their recognition is unclear. Here, we report the recognition between yeast SNARE Vti1p and its adaptor Ent3p derived from three crystal structures. Surprisingly, this yeast pair Vti1p/Ent3p interacts through a distinct binding site compared to their homologues vti1b/epsinR in mammals. An opposite surface on Vti1p_Habc domain binds to a conserved area on the epsin N-terminal homology (ENTH) domain of Ent3p. Two-hybrid, in vitro pull-down and in vivo experiments indicate this binding interface is important for correct localization of Vti1p in the cell. This previously undescribed discovery that a cargo and adaptor pair uses different binding sites across species suggests the diversity of SNARE-adaptor recognition in vesicular transport.
authors: University of Science and Technology of China, Hefei, Anhui 230026, China.
authors: Jing
authors: J
authors: Wang
authors: Michael
authors: M
authors: Gossing
authors: Pengfei
authors: P
authors: Fang
authors: Jana
authors: J
authors: Zimmermann
authors: Xu
authors: X
authors: Li
authors: Gabriele Fischer
authors: GF
authors: von Mollard
authors: Liwen
authors: L
authors: Niu
authors: Maikun
authors: M
authors: Teng
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D006801:Humans; D008561:Membrane Fusion / Q000502:physiology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D054730:Protein Interaction Domains and Motifs; D050766:Qb-SNARE Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D050600:SNARE Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C495587:Ent3 protein, S cerevisiae; D050766:Qb-SNARE Proteins; D011994:Recombinant Proteins; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C106736:VTI1 protein, S cerevisiae; C114154:epsin