ArticleBiophysical journal2011
Single vesicle assaying of SNARE-synaptotagmin-driven fusion reveals fast and slow modes of both docking and fusion and intrasample heterogeneity.
Article in Biophysical journal, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- From Population Averaging to Single Event Resolution: Evolution of Sensing Platforms for Membrane Fusion.Sensors (Basel, Switzerland) · 2026Review
- Dynamic Fusion of Nucleic Acid Functionalized Nano-/Micro-Cell-Like Containments: From Basic Concepts to Applications.ACS nano · 2023Review
- Single-particle combinatorial multiplexed liposome fusion mediated by DNA.Nature chemistry · 2022Article
- Review
- Search for a minimal machinery for CaNeuroscience · 2019Article
- MicroRNA Detection by DNA-Mediated Liposome Fusion.Chembiochem : a European journal of chemical biology · 2018Article
- Review
- Visualization and Sequencing of Membrane Remodeling Leading to Influenza Virus Fusion.Journal of virology · 2016Article
- Nonaggregated α-synuclein influences SNARE-dependent vesicle docking via membrane binding.Biochemistry · 2014Article
- Mechanism for calcium ion sensing by the C2A domain of synaptotagmin I.Biophysical journal · 2012Article
- A single vesicle-vesicle fusion assay for in vitro studies of SNAREs and accessory proteins.Nature protocols · 2012Article
- Intermembrane docking reactions are regulated by membrane curvature.Biophysical journal · 2011Article
- Mixing subattolitre volumes in a quantitative and highly parallel manner with soft matter nanofluidics.Nature nanotechnology · 2011Article
- Sensing-applications of surface-based single vesicle arrays.Sensors (Basel, Switzerland) · 2010Review
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3 authors.
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Abstract
Lipid mixing between vesicles functionalized with SNAREs and the cytosolic C2AB domain of synaptotagmin-1 recapitulates the basic Ca(2+) dependence of neuronal exocytosis. However, in the conventional ensemble lipid mixing assays it is not possible to discriminate whether Ca(2+) accelerates the docking or the fusion of vesicles. Here we report a fluorescence microscopy-based assay to monitor SNARE-mediated docking and fusion of individual vesicle pairs. In situ measurement of the concentration of diffusing particles allowed us to quantify docking rates by a maximum-likelihood approach. This analysis showed that C2AB and Ca(2+) accelerate vesicle-vesicle docking with more than two orders of magnitude. Comparison of the measured docking rates with ensemble lipid mixing kinetics, however, suggests that in most cases bilayer fusion remains the rate-limiting step. Our single vesicle results show that only ∼60% of the vesicles dock and only ∼6% of docked vesicles fuse. Lipid mixing on single vesicles was fast (t(mix) < 1 s) while an ensemble assay revealed two slow mixing processes with t(mix) ∼ 1 min and t(mix) ∼ 20 min. The presence of several distinct docking and fusion pathways cannot be rationalized at this stage but may be related to intrasample heterogeneities, presumably in the form of lipid and/or protein composition.
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