High-resolution electron cryomicroscopy of V-ATPase in native synaptic vesicles

• 成果类型: Article
• 署名作者: Coupland, Claire E.; Karimi, Ryan; Bueler, Stephanie A.; Liang, Yingke; Courbon, Gautier M.; Di Trani, Justin M.; Wong, Cassandra J.; Saghian, Rayan; Youn, Ji-Young; Wang, Lu-Yang; Rubinstein, John L.
• 署名单位: University of Toronto; Hospital for Sick Children (SickKids); University of Toronto; University of Toronto; University of Toronto; Sinai Health System Toronto; Lunenfeld Tanenbaum Research Institute; University of Toronto; Hospital for Sick Children (SickKids); University of Toronto; University of Toronto; Hospital for Sick Children (SickKids); University of Toronto
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-13836
• DOI: 10.1126/science.adp5577
• 发表日期: 2024-07-12
• 页码: 168-174
• 关键词: vacuolar h+-atpase; cryo-em; membrane-protein; plasma-membrane; synaptophysin; synaptobrevin; channel; domain; v-1; identification

摘要

Intercellular communication in the nervous system occurs through the release of neurotransmitters into the synaptic cleft between neurons. In the presynaptic neuron, the proton pumping vesicular- or vacuolar-type ATPase (V-ATPase) powers neurotransmitter loading into synaptic vesicles (SVs), with the V(1 )complex dissociating from the membrane region of the enzyme before exocytosis. We isolated SVs from rat brain using SidK, a V-ATPase-binding bacterial effector protein. Single-particle electron cryomicroscopy allowed high-resolution structure determination of V-ATPase within the native SV membrane. In the structure, regularly spaced cholesterol molecules decorate the enzyme's rotor and the abundant SV protein synaptophysin binds the complex stoichiometrically. ATP hydrolysis during vesicle loading results in a loss of the V-1 region of V-ATPase from the SV membrane, suggesting that loading is sufficient to induce dissociation of the enzyme.