title: Synaptophysin I selectively specifies the exocytic pathway of synaptobrevin 2/VAMP2.
abstract: Biogenesis and recycling of synaptic vesicles are accompanied by sorting processes that preserve the molecular composition of the compartments involved. In the present study, we have addressed the targeting of synaptobrevin 2/VAMP2 (vesicle-associated membrane protein 2), a critical component of the synaptic vesicle--fusion machinery, in a heterotypic context where its sorting is not confounded by the presence of other neuron-specific molecules. Ectopically expressed synaptophysin I interacts with VAMP2 and alters its default surface targeting to a prominent vesicular distribution, with no effect on the targeting of other membrane proteins. Protein-protein interaction is not sufficient for the control of VAMP2 sorting, which is mediated by the C-terminal domain of synaptophysin I. Synaptophysin I directs the sorting of VAMP2 to vesicles before surface delivery, without influencing VAMP2 endocytosis. Consistent with this, dynamin and alpha-SNAP (soluble N-ethylmaleimide-sensitive fusion protein-attachment protein) mutants which block trafficking at the plasma membrane do not abrogate the effect of synaptophysin I on VAMP2 sorting. These results indicate that the sorting determinants of synaptic vesicle proteins can operate independently of a neuronal context and implicate the association of VAMP2 with synaptophysin I in the specification of the pathway of synaptic vesicle biogenesis.
authors.affiliation: San Raffaele Scientific Institute and Vita-Salute University, Via Olgettina 58, 20132 Milan, Italy.
authors.forename: Dario
authors.initials: D
authors.lastname: Bonanomi
authors.forename: Laura
authors.initials: L
authors.lastname: Rusconi
authors.forename: Chiara Agnese
authors.initials: CA
authors.lastname: Colombo
authors.forename: Fabio
authors.initials: F
authors.lastname: Benfenati
authors.forename: Flavia
authors.initials: F
authors.lastname: Valtorta
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D034281:Dynamins / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D016708:Synaptophysin / Q000235:genetics / Q000378:metabolism*; D050988:Vesicle-Associated Membrane Protein 2 / Q000235:genetics / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D016708:Synaptophysin; D050988:Vesicle-Associated Membrane Protein 2; D034281:Dynamins