TULIPs: tunable, light-controlled interacting protein tags for cell biology.
TULIPs: tunable, light-controlled interacting protein tags for cell biology.
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DOI：
10.1038/nmeth.1904
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发表时间：
2012-03-04
期刊：
影响因子：
48
通讯作者：
Glotzer M
中科院分区：
文献类型：
--
作者：
Strickland D;Lin Y;Wagner E;Hope CM;Zayner J;Antoniou C;Sosnick TR;Weiss EL;Glotzer M
自然而然，光开关蛋白质提供了一种直接控制蛋白质复合体形成的方法，这种复合体驱动了多种细胞过程。我们开发了可调谐的光诱导二聚标签(郁金香)，该标签是基于燕麦向光蛋白1的LOV2结构域(AsLOV2)和工程PDZ结构域(EPDZ)之间的合成作用而开发的。郁金香标签可以在活的酵母和哺乳动物细胞中招募蛋白质到不同的结构中，无论是在全球范围内，还是通过使用可操纵的激光进行精确的空间控制。这种相互作用的平衡结合和动力学参数可以通过突变来调节，这使得郁金香很容易适应不同敏感性和反应时间的信号通路。我们展示了郁金香的用途，通过赋予酵母交配途径的功能不同的组成部分光敏感性，并通过指导细胞极化的位置。
Naturally photoswitchable proteins offer a means of directly manipulating the formation of protein complexes that drive a diversity of cellular processes. We have developed tunable light-inducible dimerization tags (TULIPs) based on a synthetic interaction between the LOV2 domain of Avena sativa phototropin 1 (AsLOV2) and an engineered PDZ domain (ePDZ). TULIP tags can recruit proteins to diverse structures in living yeast and mammalian cells, either globally or with precise spatial control using a steerable laser. The equilibrium binding and kinetic parameters of the interaction are tunable by mutation, making TULIPs readily adaptable to signaling pathways with varying sensitivities and response times. We demonstrate the utility of TULIPs by conferring light sensitivity to functionally distinct components of the yeast mating pathway and by directing the site of cell polarization.