Functional significance of human trp1 and trp3 in store-operated Ca2+ entry in HEK-293 cells
Functional significance of human trp1 and trp3 in store-operated Ca2+ entry in HEK-293 cells
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DOI：
10.1152/ajpcell.2000.278.3.c526
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发表时间：
2000-03-01
影响因子：
5.5
通讯作者：
Villereal, ML
中科院分区：
文献类型：
--
作者：
Wu, XY;Babnigg, G;Villereal, ML
果蝇的瞬时受体电位（trp）基因似乎编码果蝇的存储操作通道（SOC），一些哺乳动物的trp同源物已被提出编码哺乳动物的SOC。这项研究提供了证据的表达三个trp同源物（Mtrp 2，Mtrp 3，和Mtrp 4）在野生型和src基因敲除小鼠的成纤维细胞，和四个trp同源物（Htrp 1，Htrp 3，Htrp 4，和Htrp 6）在人胚肾（HEK-293）细胞的基础上，RT-PCR技术。在HEK-293细胞中稳定转染了323 bp的Htrp 3反义构建体（Htrp 3AS），北方印迹分析显示，与在稳定转染了短Htrp 3正义构建体（Htrp 3S）的细胞中观察到的相比，4 kb转录物的表达被显著抑制。与Htrp 3S细胞相比，Htrp 3AS细胞中的SOC活性（用毒胡萝卜素耗尽Ca 2+储存后的Ba 2+内流监测）降低了32%。在稳定表达Htrp 3AS的细胞中瞬时转染369 bp的Htrp 1反义构建体诱导了更高水平的抑制（55%）的钙池操作的Ca 2+进入。这些数据表明，Htrp 1和Htrp 3可能是SOC的功能亚基。
The Drosophila trp (transient receptor potential) gene appears to encode the Drosophila store-operated channel(SOC), and some mammalian trp homologues have been proposed to encode mammalian SOCs. This study provides evidence for the expression of three trp homologues (Mtrp2, Mtrp3, and Mtrp4) in fibroblasts from wild-type and src knockout mice, and four trp homologues (Htrp1, Htrp3, Htrp4, and Htrp6) in human embryonic kidney (HEK-293) cells based on RT-PCR techniques. In HEK-293 cells stably transfected with a 323-bp Htrp3 antisense construct (Htrp3AS), Northern blot analysis revealed that the expression of a 4-kb transcript was dramatically suppressed in comparison to that observed in cells stably transfected with a short Htrp3 sense construct (Htrp3S). Activity of SOCs, monitored as Ba2+ influx following Ca2+ store depletion with thapsigargin, was reduced by 32% in Htrp3AS cells in comparison with Htrp3S cells. Transient transfection of a 369-bp Htrp1 antisense construct in cells stably expressing Htrp3AS induced a higher level of inhibition (55%) of store-operated Ca2+ entry. These data suggest that Htrp1 and Htrp3 may be functional subunits of SOCs.