Isolation of inositol 1, 4, 5-trisphosphate receptor-associating proteins and selective knockdown using RNA interference
Isolation of inositol 1, 4, 5-trisphosphate receptor-associating proteins and selective knockdown using RNA interference
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使用 RNA 干扰分离肌醇 1,4,5-三磷酸受体相关蛋白并选择性敲低
DOI：
10.1007/978-1-60327-175-2_9
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Mikoshiba K
中科院分区：
文献类型：
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作者：
Mizutani A;Kawaai K;Hisatsune C;Ando H;Michikawa T;Mikoshiba K
三磷酸肌醇（Inositol 1，4，5-trisphosphate，IP 3）受体（IP 3Rs）是位于细胞内Ca ~（2+）库上的IP 3门控Ca ~（2+）释放通道，在产生复杂的细胞内Ca ~（2+）信号模式中发挥作用。我们在此显示了使用亲和柱层析鉴定来自各种细胞和组织的IP 3R同种型的相关蛋白以及使用RNA干扰特异性敲低IP 3R同种型及其相关蛋白的表达的实验方案。这些方法将提供线索，以了解信号复合物如何有助于细胞内Ca 2+信号的时空模式的产生的确切性质。
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.