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Studying the Effects of Inositol Pyrophosphates in an In Vitro Vesicle–Vesicle Fusion Assay

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Inositol Phosphates

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2091))

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Abstract

In a recent in vitro vesicle fusion study in which we monitored the fusion of reconstituted SNARE and synaptotagmin-containing proteoliposomes, we discovered that inositol pyrophosphate (5-IP7) is a potent inhibitor of neuronal exocytosis. We found that the inhibitory effect of 5-IP7, which is 10 times more potent than those of IP6 and 1-IP7, requires direct interaction with synaptotagmin. Here, we provide a detailed protocol for proteoliposome preparation and bulk observation of proteoliposome fusion based on FRET signals.

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Acknowledgments

This work was supported by the National Creative Research Initiative Program (Center for Single-Molecule Systems Biology to T.-Y.Y).

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Authors and Affiliations

  1. National Creative Research Center for Single-Molecule Systems Biology, School of Biological Sciences, Seoul National University, Seoul, South Korea

    Mi Sook Sung & Tae-Young Yoon

  2. Center for Theragnosis, Korea Institute of Science and Technology (KIST), Seoul, South Korea

    Mi Sook Sung & Tae-Young Yoon

Authors
  1. Mi Sook Sung
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  2. Tae-Young Yoon
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Corresponding author

Correspondence to Tae-Young Yoon .

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Editors and Affiliations

  1. Department of Chemistry, The Catholic University of America, Washington, DC, USA

    Gregory J. Miller

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Sung, M.S., Yoon, TY. (2020). Studying the Effects of Inositol Pyrophosphates in an In Vitro Vesicle–Vesicle Fusion Assay. In: Miller, G. (eds) Inositol Phosphates. Methods in Molecular Biology, vol 2091. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0167-9_13

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  • DOI: https://doi.org/10.1007/978-1-0716-0167-9_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0166-2

  • Online ISBN: 978-1-0716-0167-9

  • eBook Packages: Springer Protocols

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