Quantitative Immunofluorescence for Measuring Spatial Compartmentation of Covalently Modified Signaling Proteins
Quantitative Immunofluorescence for Measuring Spatial Compartmentation of Covalently Modified Signaling Proteins
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DOI：
10.1109/icimw.2009.5324773
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发表时间：
2009-01-01
期刊：
影响因子：
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通讯作者：
Asthagiri, Anand R.
中科院分区：
文献类型：
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作者：
Kim, Jin-Hong;Asthagiri, Anand R.
细胞内信号通路控制细胞行为和多细胞形态动力学。对这些通路的定量理解将为调节这些信号提供设计原则，以便工程化细胞行为和组织形态。信号通路中的信息传递涉及信号蛋白的位点特异性共价修饰和空间定位。在这里，我们描述了一种算法，用于定量的空间定位的共价修饰的信号蛋白从免疫荧光（IF）染色获得的图像。作为一个案例研究，我们应用该方法来定量的双重磷酸化细胞外调节激酶（ERK）在细胞核中的量。这里提出的算法提供了一个通用的示意图，可以修改和更广泛地应用于量化其他共价修饰的信号蛋白的空间区室化。
Intracellular signaling pathways control cell behaviors and multicellular morphodynamics. A quantitative understanding of these pathways will provide design principles for tuning these signals in order to engineer cell behaviors and tissue morphology. The transmission of information in signaling pathways involves both site-specific covalent modifications and spatial localization of signaling proteins. Here, we describe an algorithm for quantifying the spatial localization of covalently modified signaling proteins from images acquired by immunofluorescence (IF) staining. As a case study, we apply the method to quantify the amount of dually phosphorylated extracellular-regulated kinase (ERK) in the nucleus. The algorithm presented here provides a general schematic that can be modified and applied more broadly to quantify the spatial compartmentation of other covalently modified signaling proteins.