Interaction assays in yeast and cultured cells confirm known and identify novel partners of the synaptic vesicle protein synaptophysin

Neuroscience
M Felkl, R E Leube

Abstract

Synaptophysin (SYP) is a major protein of neurotransmitter-containing vesicles spanning the membrane four times and contributing to various aspects of the synaptic vesicle cycle. The split-ubiquitin yeast two-hybrid system was used to characterize molecular interactions of membrane-bound, full-length murine SYP. In this way, the known homophilic SYP-SYP association could be confirmed and heterophilic binding of SYP to other tetraspan vesicle membrane proteins of the secretory carrier-associated membrane- and synaptogyrin-type could be detected for the first time. SYP-binding was also observed for the vSNARE synaptobrevin2 and various membrane and membrane-associated proteins. Double labeling immunofluorescence microscopy of murine retina, co-immunoprecipitation experiments and fluorescence energy resonance transfer (FRET) analyses between fluorescent protein-tagged polypeptides were carried out to validate and further characterize the association of SYP with the tetraspan vesicle membrane proteins secretory carrier-associated membrane protein 1 and synaptogyrin3, with synaptobrevin2, and the newly identified binding partners phospholipase D4, stathmin-like3, Rho family GTPase2 and ADP-ribosylation factor interacting protein2. I...Continue Reading

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Citations

Apr 10, 2012·PloS One·Pasano Bojang, Ronald G Gregg
Aug 24, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Sarah L Gordon, Michael A Cousin
Oct 1, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Sarah L GordonMichael A Cousin
Jul 8, 2009·International Journal of Molecular Sciences·Anna BrücknerUwe Schlattner
Feb 24, 2016·Frontiers in Synaptic Neuroscience·Sarah L Gordon, Michael A Cousin
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Mar 27, 2021·The Journal of Biological Chemistry·Callista B HarperMichael A Cousin

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