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首页> 外文期刊>Cell Calcium: The International Interdisciplinary Forum for Research on Calcium >Expression of a truncated form of inositol 1,4,5-trisphosphate receptor type III in the cytosol of DT40 triple inositol 1,4,5-trisphosphate receptor-knockout cells.
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Expression of a truncated form of inositol 1,4,5-trisphosphate receptor type III in the cytosol of DT40 triple inositol 1,4,5-trisphosphate receptor-knockout cells.

机译:截断形式的肌醇1,4,5-三磷酸受体III型在DT40三重肌醇1,4,5-三磷酸受体敲除细胞的细胞质中的表达。

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摘要

In non-excitable cells, the inositol 1,4,5-trisphosphate receptor (IP(3)R) is an intracellular Ca(2+) channel playing a major role in Ca(2+) signaling. Three isoforms of IP(3)R have been identified and most cell types express different proportions of each isoform. The DT40 B lymphocyte cell line lacking all three IP(3)R isoforms (DT40IP(3)R-KO cells) represents an excellent model to re-express any recombinant IP(3)R and analyze its specific properties. In the study presented here, we confirmed that DT40IP(3)R-KO cells do not express any IP(3)-sensitive Ca(2+) release channel. However, with an immunoblot approach and a [ [Formula: see text] ]IP(3) binding approach we demonstrated the presence of a C-terminally truncated form of IP(3)R type III in the cytosolic fraction of DT40IP(3)R-KO cells. We further showed that this truncated IP(3)R retained the ability to couple to the Ca(2+) entry channel TRPC6. Therefore, a word of caution is offered about the interpretation of results obtained in using DT40IP(3)R-KO cells to study the cellular mechanisms of Ca(2+) entry.
机译:在非兴奋性细胞中,肌醇1,4,5-三磷酸受体(IP(3)R)是细胞内Ca(2+)通道,在Ca(2+)信号传导中起主要作用。 IP(3)R的三种同工型已被鉴定,大多数细胞类型表达每种同工型的不同比例。缺少所有三个IP(3)R同工型(DT40IP(3)R-KO细胞)的DT40 B淋巴细胞细胞系代表了一个很好的模型,可以重新表达任何重组IP(3)R并分析其特定特性。在这里介绍的研究中,我们确认DT40IP(3)R-KO细胞不表达任何IP(3)敏感的Ca(2+)释放通道。但是,通过免疫印迹方法和[[公式:参见文本]] IP(3)结合方法,我们证明了DT40IP(3)的胞质部分中存在IP端3型IP(3)R型C末端截短形式R-KO细胞。我们进一步表明,此截断的IP(3)R保留了耦合到Ca(2+)入口通道TRPC6的能力。因此,对于使用DT40IP(3)R-KO细胞研究Ca(2+)进入的细胞机制所获得的结果的解释,请谨慎使用。

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