Fusion pore formation and expansion induced by Ca2+ and synaptotagmin 1

Authors
Lai, YingDiao, JiajieLiu, YanxinIshitsuka, YujiSu, ZengliuSchulten, KlausHa, TaekjipShin, Yeon-Kyun
Issue Date
2013-01-22
Publisher
NATL ACAD SCIENCES
Citation
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.110, no.4, pp.1333 - 1338
Abstract
Fusion pore formation and expansion, crucial steps for neurotransmitter release and vesicle recycling in soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-dependent vesicle fusion, have not been well studied in vitro due to the lack of a reliable content-mixing fusion assay. Using methods detecting the intervesicular mixing of small and large cargoes at a single-vesicle level, we found that the neuronal SNARE complexes have the capacity to drive membrane hemifusion. However, efficient fusion pore formation and expansion require synaptotagmin 1 and Ca2+. Real-time measurements show that pore expansion detected by content mixing of large DNA cargoes occurs much slower than initial pore formation that transmits small cargoes. Slow pore expansion perhaps provides a time window for vesicles to escape the full collapse fusion pathway via alternative mechanisms such as kiss-and-run. The results also show that complexin 1 stimulates pore expansion significantly, which could put bias between two pathways of vesicle recycling.
Keywords
MEDIATED MEMBRANE-FUSION; VESICLE FUSION; MOLECULAR-DYNAMICS; CA2+-TRIGGERED EXOCYTOSIS; NEUROTRANSMITTER RELEASE; SNARE COMPLEX; IN-VITRO; HEMIFUSION; ASSAY; BINDING; MEDIATED MEMBRANE-FUSION; VESICLE FUSION; MOLECULAR-DYNAMICS; CA2+-TRIGGERED EXOCYTOSIS; NEUROTRANSMITTER RELEASE; SNARE COMPLEX; IN-VITRO; HEMIFUSION; ASSAY; BINDING; membrane fusion; synaptic vesicle; fusion pore dynamics; DNA probe; single molecule
ISSN
0027-8424
URI
https://pubs.kist.re.kr/handle/201004/128438
DOI
10.1073/pnas.1218818110
Appears in Collections:
KIST Article > 2013
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