ArticleBiophysical journal2011
Docking, not fusion, as the rate-limiting step in a SNARE-driven vesicle fusion assay.
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 17 papers.
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Who cites it
17 citing papers in PubMed.
- Optimal Detection of Fusion Pore Dynamics Using Polarized Total Internal Reflection Fluorescence Microscopy.Frontiers in molecular biosciences · 2021Article
- SNARE-Mediated Fusion of Single Chromaffin Granules with Pore-Spanning Membranes.Biophysical journal · 2019Article
- Highly Efficient Protein-free Membrane Fusion: A Giant Vesicle Study.Biophysical journal · 2019Article
- Reconstitution of Human Ion Channels into Solvent-free Lipid Bilayers Enhanced by Centrifugal Forces.Biophysical journal · 2016Article
- Article
- Variable cooperativity in SNARE-mediated membrane fusion.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Nonaggregated α-synuclein influences SNARE-dependent vesicle docking via membrane binding.Biochemistry · 2014Article
- Preparation and characterization of SNARE-containing nanodiscs and direct study of cargo release through fusion pores.Nature protocols · 2013Article
- Coiled-coil formation on lipid bilayers--implications for docking and fusion efficiency.Biophysical journal · 2012Article
- Mechanism for calcium ion sensing by the C2A domain of synaptotagmin I.Biophysical journal · 2012Article
- Solution single-vesicle assay reveals PIP2-mediated sequential actions of synaptotagmin-1 on SNAREs.The EMBO journal · 2012Article
- A single vesicle-vesicle fusion assay for in vitro studies of SNAREs and accessory proteins.Nature protocols · 2012Article
- Fusion of single proteoliposomes with planar, cushioned bilayers in microfluidic flow cells.Nature protocols · 2012Article
- Munc13-4 reconstitutes calcium-dependent SNARE-mediated membrane fusion.The Journal of cell biology · 2012Article
- SNARE proteins: one to fuse and three to keep the nascent fusion pore open.Science (New York, N.Y.) · 2012Article
- Spatial modeling of vesicle transport and the cytoskeleton: the challenge of hitting the right road.PloS one · 2012Article
- Inside insight to membrane fusion.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
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2 authors.
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Abstract
In vitro vesicle fusion assays that monitor lipid mixing between t-SNARE and v-SNARE vesicles in bulk solution exhibit remarkably slow fusion on the nonphysiological timescale of tens of minutes to several hours. Here, single-vesicle, fluorescence resonance energy transfer-based assays cleanly separate docking and fusion steps for individual vesicle pairs containing full-length SNAREs. Docking is extremely inefficient and is the rate-limiting step. Of importance, the docking and fusion kinetics are comparable in the two assays (one with v-SNARE vesicles tethered to a surface and the other with v-SNARE vesicles free in solution). Addition of the V(C) peptide synaptobrevin-2 (syb(57-92)) increases the docking efficiency by a factor of ∼30, but docking remains rate-limiting. In the presence of V(C) peptide, the fusion step occurs on a timescale of ∼10 s. In previous experiments involving bulk fusion assays in which the addition of synaptotagmin/Ca(2+), Munc-18, or complexin accelerated the observed lipid-mixing rate, the enhancement may have arisen from the docking step rather than the fusion step.
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