Lai, Ying Diao, Jiajie Liu, Yanxin Ishitsuka, Yuji Su, Zengliu Schulten, Klaus Ha, Taekjip Shin, Yeon-Kyun 2024-01-20T13:03:22Z 2024-01-20T13:03:22Z 2021-09-01 2013-01-22 0027-8424 https://pubs.kist.re.kr/handle/201004/128438 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. English NATL ACAD SCIENCES MEDIATED MEMBRANE-FUSION VESICLE FUSION MOLECULAR-DYNAMICS CA2+-TRIGGERED EXOCYTOSIS NEUROTRANSMITTER RELEASE SNARE COMPLEX IN-VITRO HEMIFUSION ASSAY BINDING Fusion pore formation and expansion induced by Ca2+ and synaptotagmin 1 Article 10.1073/pnas.1218818110 1 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.110, no.4, pp.1333 - 1338 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 110 4 1333 1338 scie scopus 000314453900044 2-s2.0-84872850714 Multidisciplinary Sciences Science & Technology - Other Topics Article 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