Purified TPC isoforms form NAADP receptors with distinct roles for Ca(2+) signaling and endolysosomal trafficking.
Purified TPC isoforms form NAADP receptors with distinct roles for Ca(2+) signaling and endolysosomal trafficking.
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DOI：
10.1016/j.cub.2010.02.049
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发表时间：
2010-04-27
期刊：
影响因子：
9.2
通讯作者：
Galione, Antony
中科院分区：
文献类型：
--
作者：
Ruas, Margarida;Rietdorf, Katja;Arredouani, Abdelilah;Davis, Lianne C.;Lloyd-Evans, Emyr;Koegel, Heidi;Funnell, Timothy M.;Morgan, Anthony J.;Ward, John A.;Watanabe, Keiko;Cheng, Xiaotong;Churchill, Grant C.;Zhu, Michael X.;Platt, Frances M.;Wessel, Gary M.;Parrington, John;Galione, Antony
细胞内Ca ~（2+）信号是细胞信号转导的关键环节。在所描述的三种主要的Ca 2+动员信使中，最有效的烟酸腺嘌呤二核苷酸磷酸（NAADP）在其分子靶点方面是最不清楚的。最近，我们发现，异源表达的双孔通道（TPC）蛋白增强NAADP诱导的Ca 2+释放，而NAADP的反应被取消在胰腺β细胞Tpcn 2基因敲除小鼠。然而，TPC是否构成天然NAADP受体尚不清楚。在这里，我们表明，免疫纯化的内源性TPC复合物具有归因于NAADP受体的标志性特性，包括纳摩尔配体亲和力。我们的研究还揭示了三种TPC亚型之间的重要功能差异。因此，TPC 1和TPC 2都介导NAADP诱导的Ca 2+释放，但随后IP 3Rs对该触发Ca 2+的放大与TPC 2更紧密地耦合。相反，TPC 3表达抑制NAADP诱导的Ca 2+释放。最后，TPC表达增加对内溶酶体结构和动力学有显著和对比的影响，暗示NAADP在调节囊泡运输中的作用。我们建议，NAADP调节内溶酶体Ca 2+的存储和释放通过TPC和协调内质网Ca 2+释放的作用，影响Ca 2+信号在健康和疾病。●内源性TPC蛋白复合物具有天然NAADP受体的标志性特性● TPC差异定位于NAADP诱导的Ca 2+释放的亚细胞位点●不同的TPC亚型在NAADP诱导的Ca 2+释放中显示出不同的作用●改变的TPC表达对内溶酶体功能具有显著的差异性影响
Intracellular Ca2+ signals constitute key elements in signal transduction. Of the three major Ca2+ mobilizing messengers described, the most potent, nicotinic acid adenine dinucleotide phosphate (NAADP) is the least well understood in terms of its molecular targets. Recently, we showed that heterologous expression of two-pore channel (TPC) proteins enhances NAADP-induced Ca2+ release, whereas the NAADP response was abolished in pancreatic beta cells from Tpcn2 gene knockout mice. However, whether TPCs constitute native NAADP receptors is unclear. Here we show that immunopurified endogenous TPC complexes possess the hallmark properties ascribed to NAADP receptors, including nanomolar ligand affinity. Our study also reveals important functional differences between the three TPC isoforms. Thus, TPC1 and TPC2 both mediate NAADP-induced Ca2+ release, but the subsequent amplification of this trigger Ca2+ by IP3Rs is more tightly coupled for TPC2. In contrast, TPC3 expression suppressed NAADP-induced Ca2+ release. Finally, increased TPC expression has dramatic and contrasting effects on endolysosomal structures and dynamics, implicating a role for NAADP in the regulation of vesicular trafficking. We propose that NAADP regulates endolysosomal Ca2+ storage and release via TPCs and coordinates endoplasmic reticulum Ca2+ release in a role that impacts on Ca2+ signaling in health and disease. ► Endogenous TPC protein complex has hallmark properties of native NAADP receptors ► TPCs differentially localize to subcellular sites of NAADP-induced Ca2+ release ► Different TPC isoforms show differential roles in NAADP-evoked Ca2+ release ► Altered TPC expression has dramatic, differential effects on endolysosomal function
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DOI：
10.1006/bbrc.2000.3444
发表时间：
2000-09-16
影响因子：
3.1
作者：
Billington, RA;Genazzani, AA
通讯作者：
Genazzani, AA
影响因子：
3.9
作者：
Galione, A.
通讯作者：
Galione, A.
影响因子：
4
作者：
Parkesh, Raman;Lewis, Alexander M.;Churchill, Grant C.
通讯作者：
Churchill, Grant C.
DOI：
10.1083/jcb.200904073
发表时间：
2009-07-27
期刊：
The Journal of cell biology
影响因子：
--
作者：
Brailoiu E;Churamani D;Cai X;Schrlau MG;Brailoiu GC;Gao X;Hooper R;Boulware MJ;Dun NJ;Marchant JS;Patel S
通讯作者：
Patel S
影响因子：
4.8
作者：
LEE, HC;AARHUS, R
通讯作者：
AARHUS, R