The V Sector of the V-ATPase, Synaptobrevin, and Synaptophysin Are Associated on Synaptic Vesicles in a Triton X-100-resistant, Freeze-thawing Sensitive, Complex
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Implication
Biochemical analysis reveals a freeze-thaw-sensitive complex of V-ATPase, synaptobrevin, and synaptophysin in rat synaptosomes, suggesting a regulatory mechanism prior to SNARE complex assembly.
Key Points
- To identify and characterize molecular interactions between the V0 sector of the vacuolar proton pump, synaptobrevin 2, and synaptophysin on synaptic vesicles.
- Performed immunoprecipitation on freshly prepared and freeze-thawed Triton X-100 extracts from rat synaptosomes using anti-synaptobrevin 2 and anti-synaptophysin antibodies.
- Identified interacting proteins using peptide sequencing, Western blotting, and rat brain subcellular fractionation.
- Fresh synaptosome extracts demonstrated coprecipitation of V-ATPase V0 subunits (subunit c/ductin, Ac39, and Ac116) alongside synaptobrevin 2 and synaptophysin.
- Freeze-thawing dissociated the V-ATPase subunits and synaptophysin from synaptobrevin 2, shifting association toward SNAP-25 and syntaxin 1.
- Subcellular fractionation confirmed the colocalization of V-ATPase subunit c, synaptobrevin 2, and synaptophysin within synaptic vesicles.
Cite This Study
Galli et al. (1996) studied this question.