The Use of FRET/FLIM to Study Proteins Interacting with Plant Receptor Kinases.

The Use of FRET/FLIM to Study Proteins Interacting with Plant Receptor Kinases.
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使用 FRET/FLIM 研究与植物受体激酶相互作用的蛋白质。

DOI:
10.1007/978-1-4939-7063-6_16
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发表时间:
2017
影响因子:
--
通讯作者:
Yvonne Stahl
Yvonne Stahl
中科院分区:
--
文献类型:
--
作者:
S. Weidtkamp‐Peters;Yvonne Stahl

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近年来,植物活体组织中蛋白质相互作用的研究变得越来越重要。需要用于观察体内蛋白质相互作用的高空间和时间分辨率,例如,以跟踪植物受体激酶相互作用和复合物形成随时间的变化。体内荧光或Förster共振能量转移(FRET)测量允许在亚细胞水平上详细分析自然环境中相互作用的蛋白质。特别是FRET FLIM(荧光寿命成像显微镜)测量提供了一个非常强大和可靠的工具,满足需求的研究在体内蛋白质相互作用的定量和高精度。在这里,我们将详细描述如何实际执行在体内FRET测量的受体激酶在植物中,并讨论潜在的陷阱和考虑点。
The investigation of protein interactions in living plant tissue has become of increasing importance in recent years. A high spatial and temporal resolution for the observation of in vivo protein interaction is needed, e.g., in order to follow changes of plant receptor kinase interactions and complex formation over time. In vivo fluorescence or Förster resonance energy transfer (FRET) measurements allow for detailed analyses of interacting proteins in their natural environment at a subcellular level. Especially FRET-FLIM (fluorescence lifetime imaging microscopy) measurements provide an extremely powerful and reliable tool meeting the demands for investigating in vivo protein interaction quantitatively and with high precision. Here, we will describe in detail how to practically perform in vivo FRET measurements of receptor kinases in plants and discuss potential pitfalls and points of consideration.
DOI: 10.1016/j.cub.2013.01.045
发表时间: 2013-03-04
期刊: CURRENT BIOLOGY
影响因子: 9.2
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