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- W2074883211 abstract "Single vesicle fluorescence assay detects the fluorescence signals from surface-immobilized nano-scale vesicles (typically diameter of 50 nm). There are three principal labeling positions: vesicle membranes, luminal contents and membrane proteins, each of which allows for study of different aspects of membrane-related biological processes. During the past five years, we have reported two realizations of these possibilities: Measuring the kinetics of single vesicle-vesicle fusion by labeling vesicle membranes [1-3] and detecting fusion pore opening in such single vesicle fusion by encapsulating fluorescently-labeled DNA hairpins inside vesicles [4]. In this work, by using fluorescently labeled SNARE proteins, we report on the kinetics of SNARE complex formation observed at the single-molecule level. For this purpose, we have developed an advanced single-molecule FRET technique, in which we track not one, but up to 10 proteins at the same time while keeping the precision at the single-molecule level. The measured kinetics of SNARE complex formation shows strong, positive cooperativity. We finally discuss whether we can make two single-molecule measurements in one experimental setting, for example, detecting the moment of fusion pore opening while tracking formation of multiple SNARE complexes. Such experiment would reveal quantitative correlation between multimeric structure of SNARE proteins and its functional effect on fusion pore opening.[1] Tae-Young Yoon, Burak Okumus, Fan Zhang, Yeon-Kyun Shin and Taekjip Ha. PNAS 103, 19731 (2006).[2] Tae-Young Yoon, Xiaobing Lu, Jiajie Diao, Taekjip Ha and Yeon-Kyun Shin. Nat. Struct. Mol. Biol. 15, 707 (2008).[3] Han-Ki Lee, Yoosoo Yang, Changbong Hyeon, Tae-Sun Lee, Hong-Won Lee, Dae-Hyuk Kweon, Yeon-Kyun Shin and Tae-Young Yoon. Science 328, 760 (2010).[4] Jiajie Diao, Zengliu Su, Yuji Ishitsuka, Bin Lu, Kyung Suk Lee, Ying Lai, Yeon-Kyun Shin and Taekjip Ha. Nature Communications 1, 54 (2010)." @default.
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- W2074883211 date "2011-02-01" @default.
- W2074883211 modified "2023-09-29" @default.
- W2074883211 title "Strong, Positive Cooperativity of SNARES For Fusion Pore Opening Studied At the Single-Molecule Level" @default.
- W2074883211 doi "https://doi.org/10.1016/j.bpj.2010.12.1233" @default.
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