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Biophys J. 2008 Feb 1;94(3):1084-93. Epub 2007 Oct 5.

Munc-18-1 regulates the initial release rate of exocytosis.

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The Physiological Laboratory, School of Biomedical Sciences, University of Liverpool, Crown Street, Liverpool, L69 3BX, United Kingdom.


Carbon fiber amperometry is a popular method for measuring single exocytotic events; however, the functional interpretation of the data can prove hazardous. For example, changes to vesicle transmitter levels can appear to cause changes in the timing and rate of the fusion process itself. Use of an analytical technique based on differentiation revealed that an increase in dense-core granule catecholamine content by exogenous application of l-DOPA did not affect initial release rates. Changes to the timing and amplitude of amperometric spikes from l-DOPA-treated cells are, then, likely a reflection of the increased quantal size rather than any direct effect on exocytosis itself. Applying this new analysis to individual fusion events from cells expressing Munc-18-1 with various specific point mutations demonstrated that Munc-18-1 functions at a late stage involved in the determination of the initial rate of fusion. Furthermore, a mutation of the protein that inhibits its biochemical interaction with the t-SNARE syntaxin-1 in a closed conformation caused premature termination of the fusion event. Through these two late-stage functions, Munc-18-1 could act as a key protein involved in the presynaptic control of signaling strength and duration.

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