Abstract
Inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) are IP3-gated Ca2+ release channels localized on intracellular Ca2+ stores and play a role in the generation of complex patterns of intracellular Ca2+ signals. We show herein experimental protocols for the identification of associating proteins of IP3R isoforms from various cells and tissues using affinity column chromatography and for the specific knockdown of the expression of IP3R isoforms and their associating proteins using RNA interference. These methods will provide clues to understand the exact nature of how the signaling complex contributes to the generation of spatio-temporal patterns of intracellular Ca2+ signals.
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This work was supported by grants from the Education, Culture, Sports, and Technology of Japan.
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Mizutani, A., Kawaai, K., Hisatsune, C., Ando, H., Michikawa, T., Mikoshiba, K. (2010). Isolation of Inositol 1,4,5-Trisphosphate Receptor-Associating Proteins and Selective Knockdown Using RNA Interference. In: Barker, C. (eds) Inositol Phosphates and Lipids. Methods in Molecular Biology, vol 645. Humana Press. https://doi.org/10.1007/978-1-60327-175-2_9
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DOI: https://doi.org/10.1007/978-1-60327-175-2_9
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