Visualisation and identification of the interaction between STIM1s in resting cells.
Visualisation and identification of the interaction between STIM1s in resting cells.
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可视化和识别静息细胞中刺激之间的相互作用。
DOI：
10.1371/journal.pone.0033377
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Li H
中科院分区：
文献类型：
--
作者：
He J;Yu T;Pan J;Li H
钙池操纵的钙通道是非兴奋性细胞中主要的钙内流途径，它驱动着细胞的各种基本功能。最近，STIM 1和奥赖蛋白已被鉴定为Ca 2+释放激活的Ca 2+（CRAC）通道的主要分子组分。作为CRAC通道的关键亚基，STIM 1是ER Ca 2+传感器，对于Orai 1的募集和激活至关重要。但STIM 1向Orai 1传递信息的机制仍需进一步研究。双分子荧光互补（BiFC）是研究和可视化活细胞中蛋白质-蛋白质相互作用的最先进和最强大的工具之一。我们利用BiFC和受体光漂白荧光共振能量转移（FRET）实验来可视化和确定STIM 1在静息状态下的活细胞中的状态。我们的研究结果表明，STIM 1在静息细胞中以寡聚体形式存在，而不是SAM基序，它是STIM 1的C-末端（残基233-474）是STIM 1之间相互作用的关键结构域。STIM 1寡聚体（BiFC-STIM 1）和野生型STIM 1共定位，并且在静息条件下具有纤维状分布。ER Ca 2+商店的耗尽诱导BiFC-STIM 1分布成为点状，2-APB可以预防或逆转的效果。Ca 2+储存耗尽后，BiFC-STIM 1能够在质膜附近形成与野生型STIM 1或Orai 1共定位的斑点。我们的数据还表明，BiFC-STIM 1的功能与野生型STIM 1相比没有改变。
Store-operated Ca2+ channels are a major Ca2+ entry pathway in nonexcitable cells, which drive various essential cellular functions. Recently, STIM1 and Orai proteins have been identified as the major molecular components of the Ca2+ release-activated Ca2+ (CRAC) channel. As the key subunit of the CRAC channel, STIM1 is the ER Ca2+ sensor and is essential for the recruitment and activation of Orai1. However, the mechanisms in transmission of information of STIM1 to Orai1 still need further investigation. Bimolecular fluorescence complementation (BiFC) is one of the most advanced and powerful tools for studying and visualising protein-protein interactions in living cells. We utilised BiFC and acceptor photobleaching fluorescence resonance energy transfer (FRET) experiments to visualise and determine the state of STIM1 in the living cells in resting state. Our results demonstrate that STIM1 exists in an oligomeric form in resting cells and that rather than the SAM motif, it is the C-terminus (residues 233–474) of STIM1 that is the key domain for the interaction between STIM1s. The STIM1 oligomers (BiFC-STIM1) and wild-type STIM1 colocalised and had a fibrillar distribution in resting conditions. Depletion of ER Ca2+ stores induced BiFC-STIM1 distribution to become punctate, an effect that could be prevented or reversed by 2-APB. After depletion of the Ca2+ stores, BiFC-STIM1 has the ability to form puncta that colocalise with wild-type STIM1 or Orai1 near the plasma membrane. Our data also indicate that the function of BiFC-STIM1 was not altered compared with that of wild-type STIM1.
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影响因子：
64.5
作者：
Park CY;Hoover PJ;Mullins FM;Bachhawat P;Covington ED;Raunser S;Walz T;Garcia KC;Dolmetsch RE;Lewis RS
通讯作者：
Lewis RS
影响因子：
64.8
作者：
Prakriya, Murali;Feske, Stefan;Hogan, Patrick G.
通讯作者：
Hogan, Patrick G.
影响因子：
64.8
作者：
Yeromin, Andriy V.;Zhang, Shenyuan L.;Cahalan, Michael D.
通讯作者：
Cahalan, Michael D.
DOI：
10.1073/pnas.0608358103
发表时间：
2006-11-07
影响因子：
11.1
作者：
Baba, Yoshihiro;Hayashit, Kenji;Kurosaki, Tomohiro
通讯作者：
Kurosaki, Tomohiro
影响因子：
21.3
作者：
Srikanth, Sonal;Jung, Hea-Jin;Kim, Kyun-Do;Souda, Puneet;Whitelegge, Julian;Gwack, Yousang
通讯作者：
Gwack, Yousang