在表达FLAG标签和无标签磷酸模拟心肌肌钙蛋白I的心肌细胞中的二次磷酸化。
Secondary phosphorylation in myocytes expressing FLAG-tagged and non-tagged phospho-mimetic cardiac troponin I.
作者信息
Lang Sarah E, Stevenson Tamara K, Schatz Tabea M, Biesiadecki Brandon J, Westfall Margaret V
机构信息
Department of Cardiac Surgery, University of Michigan, Ann Arbor, MI 48109, USA.
Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, MI 48109, USA.
出版信息
Data Brief. 2017 Oct 5;15:562-566. doi: 10.1016/j.dib.2017.09.066. eCollection 2017 Dec.
Secondary phosphorylation develops in myocytes expressing phospho-mimetic cardiac troponin I (cTnI) but it is not known whether multiple substitutions (e.g. cTnISDTD and cTnIS4D) cause preferential phosphorylation of the remaining endogenous or the phospho-mimetic cTnI in intact myocytes. Western analysis was performed to determine whether the FLAG/total cTnI ratios are similar for phosphorylated versus total cTnI in myocytes expressing phospho-mimetic cTnI with Asp(D) substitutions at S43/45 plus S23/24 (cTnIS4D) or T144 (cTnISDTD). Representative Western analysis of phosphorylated S23/24 (p-S23/24) and S150 (p-S150) are presented along with re-probes using an antibody which detects all cTnI (MAB1691 Ab). The level of p-S150 also is compared to results obtained using single S43D and/or S45D phospho-mimetic substitutions. These results are discussed in more detail in Lang et al. [1].
继发性磷酸化在表达磷酸化模拟心肌肌钙蛋白I（cTnI）的心肌细胞中出现，但尚不清楚多个位点的替换（如cTnISDTD和cTnIS4D）是否会导致完整心肌细胞中剩余内源性或磷酸化模拟cTnI的优先磷酸化。进行蛋白质免疫印迹分析以确定在S43/45和S23/24（cTnIS4D）或T144（cTnISDTD）处具有天冬氨酸（D）替换的磷酸化模拟cTnI的心肌细胞中，磷酸化cTnI与总cTnI的FLAG/总cTnI比率是否相似。给出了磷酸化的S23/24（p-S23/24）和S150（p-S150）的代表性蛋白质免疫印迹分析结果，以及使用检测所有cTnI的抗体（MAB1691抗体）进行的再检测结果。还将p-S150的水平与使用单个S43D和/或S45D磷酸化模拟替换获得的结果进行了比较。这些结果在Lang等人的文献[1]中有更详细的讨论。
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