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복잡계 자기조립 연구단
복잡계 자기조립 연구단
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High Affinity Host-Guest FRET Pair for Single-Vesicle Content-Mixing Assay: Observation of Flickering Fusion Events

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Title
High Affinity Host-Guest FRET Pair for Single-Vesicle Content-Mixing Assay: Observation of Flickering Fusion Events
Author(s)
Bokyoung Gong; Choi B.-K.; Kim J.-Y.; Dinesh Shetty; Young Ho Ko; Narayanan Selvapalam; Nam Ki Lee; Kimoon Kim
Publication Date
2015-07
Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, v.137, no.28, pp.8908 - 8911
Publisher
AMER CHEMICAL SOC
Abstract
Fluorescence-based single-vesicle fusion assays provide a powerful method for studying mechanisms underlying complex biological processes of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)-mediated vesicle fusion and neurotransmitter release. A crucial element of these assays is the ability of the fluorescent probe(s) to reliably detect key intermediate events of fusion pore opening and content release/mixing. Here, we report a new, reliable, and efficient single-vesicle content-mixing assay using a high affinity, fluorophore tagged host-guest pair, cucurbit[7]uril-Cy3 and adamantane-Cy5 as a fluorescence resonance energy transfer (FRET) pair. The power of these probes is demonstrated by the first successful observation of flickering dynamics of the fusion pore by in vitro assay using neuronal SNARE-reconstituted vesicles. © 2015 American Chemical Society
URI
https://pr.ibs.re.kr/handle/8788114/1990
ISSN
0002-7863
Appears in Collections:
Center for Self-assembly and Complexity(복잡계 자기조립 연구단) > Journal Papers (저널논문)
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