Molecular architecture of synaptic vesicles.

Autor:	Kravčenko U; In situ Structural Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin 13125, Germany.; Department of Biology, Humboldt University of Berlin, Berlin, Germany., Ruwolt M; Leibniz Research Institute for Molecular Pharmacology, Berlin, Germany., Kroll J; Structural Biology of Membrane-Associated Processes, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany.; Institute of Neurophysiology, Charité-Universitätsmedizin Berlin, Berlin, Germany., Yushkevich A; In situ Structural Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin 13125, Germany.; Department of Physics, Humboldt University of Berlin, Berlin, Germany., Zenkner M; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany., Ruta J; In situ Structural Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin 13125, Germany.; Leibniz Research Institute for Molecular Pharmacology, Berlin, Germany.; Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany., Lotfy R; In situ Structural Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin 13125, Germany.; Institute of Pharmacy, Free University of Berlin, Berlin, Germany., Wanker EE; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany., Rosenmund C; Institute of Neurophysiology, Charité-Universitätsmedizin Berlin, Berlin, Germany., Liu F; Leibniz Research Institute for Molecular Pharmacology, Berlin, Germany.; Charité-Universitätsmedizin Berlin, Berlin, Germany., Kudryashev M; In situ Structural Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin 13125, Germany.; Institute of Medical Physics and Biophysics, Charité-Universitätsmedizin Berlin, Berlin, Germany.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2024 Dec 03; Vol. 121 (49), pp. e2407375121. Date of Electronic Publication: 2024 Nov 27.
DOI:	10.1073/pnas.2407375121
Abstrakt:	Synaptic vesicles (SVs) store and transport neurotransmitters to the presynaptic active zone for release by exocytosis. After release, SV proteins and excess membrane are recycled via endocytosis, and new SVs can be formed in a clathrin-dependent manner. This process maintains complex molecular composition of SVs through multiple recycling rounds. Previous studies explored the molecular composition of SVs through proteomic analysis and fluorescent microscopy, proposing a model for an average SV (1). However, the structural heterogeneity and molecular architecture of individual SVs are not well described. Here, we used cryoelectron tomography to visualize molecular details of SVs isolated from mouse brains and inside cultured neurons. We describe several classes of small proteins on the SV surface and long proteinaceous densities inside SVs. We identified V-ATPases, determined a structure using subtomogram averaging, and showed them forming a complex with the membrane-embedded protein synaptophysin (Syp). Our bioluminescence assay revealed pairwise interactions between vesicle-associated membrane protein 2 and Syp and V-ATPase Voe1 domains. Interestingly, V-ATPases were randomly distributed on the surface of SVs irrespective of vesicle size. A subpopulation of isolated vesicles and vesicles inside neurons contained a partially assembled clathrin coat with an icosahedral symmetry. We observed V-ATPases under clathrin cages in several isolated clathrin-coated vesicles (CCVs). Additionally, from isolated SV preparations and within hippocampal neurons we identified clathrin baskets without vesicles. We determined their and CCVs preferential location in proximity to the cell membrane. Our analysis advances the understanding of individual SVs' diversity and their molecular architecture. Competing Interests: Competing interests statement:The authors declare no competing interest.
Databáze:	MEDLINE
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