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Investigating the InsP3 Receptor in Living Cells by Caged InsP3

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Inositol Phosphates

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

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Abstract

The inositol 1,4,5-trisphosphate receptor (InsP3R) operates as an intracellular ligand-gated Ca2+ channel and plays a pivotal role in cellular Ca2+ homeostasis across all living cells. It is activated following membrane receptor–ligand interactions and stimulation of subsequent signaling cascades involving the enzymatic breakdown of the membrane lipid phosphatidyl-4,5-bisphosphate (PIP2) into the membrane-delimited second messenger diacylglycerol (DAG) and the diffusible second messenger inositol-1,4,5-trisphophate (InsP3). Modulation of InsP3R’s activity is thus involved in a plethora of physiological and pathological processes. Here we combine membrane permeable photoactive caged-InsP3 with Ca2+ imaging techniques in living cells to study the channel’s in vivo properties. Using UV-flashes of variable energy, the activity properties of InsP3R can be investigated in great detail in its native environment.

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Authors and Affiliations

  1. Medical Faculty, Centre for Molecular Signalling (PZMS), Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, Saarland University, Homburg/Saar, Germany

    Xin Hui & Peter Lipp

Authors
  1. Xin Hui
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  2. Peter Lipp
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Corresponding author

Correspondence to Peter Lipp .

Editor information

Editors and Affiliations

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

    Gregory J. Miller

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Hui, X., Lipp, P. (2020). Investigating the InsP3 Receptor in Living Cells by Caged InsP3. 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_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0167-9_10

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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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