title: Structural insights into the Ca2+ and PI(4,5)P2 binding modes of the C2 domains of rabphilin 3A and synaptotagmin 1.
abstract: Proteins containing C2 domains are the sensors for Ca(2+) and PI(4,5)P2 in a myriad of secretory pathways. Here, the use of a free-mounting system has enabled us to capture an intermediate state of Ca(2+) binding to the C2A domain of rabphilin 3A that suggests a different mechanism of ion interaction. We have also determined the structure of this domain in complex with PI(4,5)P2 and IP3 at resolutions of 1.75 and 1.9 Å, respectively, unveiling that the polybasic cluster formed by strands β3-β4 is involved in the interaction with the phosphoinositides. A comparative study demonstrates that the C2A domain is highly specific for PI(4,5)P2/PI(3,4,5)P3, whereas the C2B domain cannot discriminate among any of the diphosphorylated forms. Structural comparisons between C2A domains of rabphilin 3A and synaptotagmin 1 indicated the presence of a key glutamic residue in the polybasic cluster of synaptotagmin 1 that abolishes the interaction with PI(4,5)P2. Together, these results provide a structural explanation for the ability of different C2 domains to pull plasma and vesicle membranes close together in a Ca(2+)-dependent manner and reveal how this family of proteins can use subtle structural changes to modulate their sensitivity and specificity to various cellular signals.
authors.affiliation: Departamento de Bioquímica y Biología Molecular A, Facultad de Veterinaria, Regional Campus of International Excellence "Campus Mare Nostrum," Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: Jaime
authors.initials: J
authors.lastname: Guillén
authors.forename: Cristina
authors.initials: C
authors.lastname: Ferrer-Orta
authors.forename: Mònica
authors.initials: M
authors.lastname: Buxaderas
authors.forename: Dolores
authors.initials: D
authors.lastname: Pérez-Sánchez
authors.forename: Marta
authors.initials: M
authors.lastname: Guerrero-Valero
authors.forename: Ginés
authors.initials: G
authors.lastname: Luengo-Gil
authors.forename: Joan
authors.initials: J
authors.lastname: Pous
authors.forename: Pablo
authors.initials: P
authors.lastname: Guerra
authors.forename: Juan C
authors.initials: JC
authors.lastname: Gómez-Fernández
authors.forename: Nuria
authors.initials: N
authors.lastname: Verdaguer
authors.forename: Senena
authors.initials: S
authors.lastname: Corbalán-García
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018360:Crystallography, X-Ray; D006801:Humans; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D050863:Synaptotagmin I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000097624:Rabphilin-3A
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002118:Calcium; D009419:Nerve Tissue Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D050863:Synaptotagmin I; D033921:Vesicular Transport Proteins; D000097624:Rabphilin-3A
keywords: PIP2; calcium; vesicle fusion