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  Rapid structural change in synaptosomal-associated protein 25 (SNAP25) precedes the fusion of single vesicles with the plasma membrane in live chromaffin cells.

Zhao, Y., Fang, Q., Herbst, A. D., Berberian, K. N., Almers, W., & Lindau, M. (2013). Rapid structural change in synaptosomal-associated protein 25 (SNAP25) precedes the fusion of single vesicles with the plasma membrane in live chromaffin cells. Proceedings of the National Academy of Sciences of the United States of America, 110(35), 14249-14254. doi:10.1073/pnas.1306699110.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-9FA4-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-80DD-2
Genre: Journal Article

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 Creators:
Zhao, Y.1, Author              
Fang, Q.1, Author              
Herbst, A. D., Author
Berberian, K. N., Author
Almers, W., Author
Lindau, M.1, Author              
Affiliations:
1Research Group of Nanoscale Cell Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1832294              

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Free keywords: TIR-FRET imaging; electrochemical imaging; time superresolution microscopy; image analysis; transmitter release
 Abstract: The SNARE complex consists of the three proteins synaptobrevin-2, syntaxin, and synaptosomal-associated protein 25 (SNAP25) and is thought to execute a large conformational change as it drives membrane fusion and exocytosis. The relation between changes in the SNARE complex and fusion pore opening is, however, still unknown. We report here a direct measurement relating a change in the SNARE complex to vesicle fusion on the millisecond time scale. In individual chromaffin cells, we tracked conformational changes in SNAP25 by total internal reflection fluorescence resonance energy transfer (FRET) microscopy while exocytotic catecholamine release from single vesicles was simultaneously recorded using a microfabricated electrochemical detector array. A local rapid and transient FRET change occurred precisely where individual vesicles released catecholamine. To overcome the low time resolution of the imaging frames needed to collect sufficient signal intensity, a method named event correlation microscopy was developed, which revealed that the FRET change was abrupt and preceded the opening of an exocytotic fusion pore by ∼90 ms. The FRET change correlated temporally with the opening of the fusion pore and not with its dilation.

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Language(s): eng - English
 Dates: 2013-08-122013-08-27
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1073/pnas.1306699110
 Degree: -

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Title: Proceedings of the National Academy of Sciences of the United States of America
Source Genre: Journal
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Pages: - Volume / Issue: 110 (35) Sequence Number: - Start / End Page: 14249 - 14254 Identifier: -