Targeted Analysis of Phosphotyrosine Signaling by Multiple Reaction Monitoring Mass Spectrometry.
Targeted Analysis of Phosphotyrosine Signaling by Multiple Reaction Monitoring Mass Spectrometry.
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通过多重反应监测质谱法对磷酸酪氨酸信号进行靶向分析。
DOI：
10.1007/978-1-4939-7154-1_17
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Payne LS
中科院分区：
文献类型：
--
作者：
Payne LS
磷酸化蛋白质组学是公正研究信号网络激活的重要工具，特别适用于揭示驱动癌症进展的异常信号传导。然而，由于基于发现的蛋白质组工作流程固有的局限性以及基于免疫亲和的替代方法所需的高质量抗体的可用性有限，在多种实验条件下验证特定磷酸位点的行为仍然具有挑战性。靶向磷酸蛋白质组学能够在多种实验条件下对特定磷酸位点进行可重复的定量。重要的是，可以根据发现蛋白质组实验中获得的数据快速设计靶向磷酸蛋白质组测定，并规避免疫亲和技术对针对特定的、潜在免疫原性较差的磷酸肽产生可靠抗体的要求。在以下协议中，我们提出了一种在多个实验条件和/或技术和生物复制中对磷酸位点进行相对定量的方法。
Phosphoproteomics is an important tool for the unbiased investigation of signaling network activation and has particular application to unraveling aberrant signaling driving cancer progression. However, validating the behavior of specific phosphosites across multiple experimental conditions remains challenging, due to limitations inherent in discovery-based proteomic workflows and the limited availability of high-quality antibodies required for alternative, immunoaffinity-based methods. Targeted phosphoproteomics enables specific phosphosites to be quantified reproducibly across multiple experimental conditions. Importantly, targeted phosphoproteomic assays can be designed rapidly on the basis of data acquired in discovery proteomic experiments and circumvent the requirement of immunoaffinity techniques for reliable antibodies raised to specific, potentially poorly immunogenic phosphopeptides. In the following protocol, we present a method for the relative quantification of phosphosites across multiple experimental conditions and/or technical and biological replicates.
影响因子：
4.4
作者：
Tabb, David L.;Vega-Montoto, Lorenzo;Rudnick, Paul A.;Variyath, Asokan Mulayath;Ham, Amy-Joan L.;Bunk, David M.;Kilpatrick, Lisa E.;Billheimer, Dean D.;Blackman, Ronald K.;Cardasis, Helene L.;Carr, Steven A.;Clauser, Karl R.;Jaffe, Jacob D.;Kowalski, Kevin A.;Neubert, Thomas A.;Regnier, Fred E.;Schilling, Birgit;Tegeler, Tony J.;Wang, Mu;Wang, Pei;Whiteaker, Jeffrey R.;Zimmerman, Lisa J.;Fisher, Susan J.;Gibson, Bradford W.;Kinsinger, Christopher R.;Mesri, Mehdi;Rodriguez, Henry;Stein, Stephen E.;Tempst, Paul;Paulovich, Amanda G.;Liebler, Daniel C.;Spiegelman, Cliff
通讯作者：
Spiegelman, Cliff
影响因子：
4.8
作者：
Colangelo, Christopher M.;Chung, Lisa;Bruce, Can;Cheung, Kei-Hoi
通讯作者：
Cheung, Kei-Hoi