Peng Yuanliang, Tang Li, Li Yanan, Song Jianhui, Liu Hong, Wang Pan, Zhong Zhizhou, Yang Yifei, Wang Shihui, Chen Lixiang, Zhang Ji, Zhang Shijie, Wang Zi, Li Min, Liang Long, Liu Jing
Department of Hematology, The Second Xiangya Hospital of Central South University, Molecular Biology Research Center, Center for Medical Genetics, School of Life Sciences, Hunan Province Key Laboratory of Basic and Applied Hematology, Central South University, Changsha, China.
School of Information Science and Engineering, Central South University, Changsha, China.
Br J Haematol. 2022 Nov;199(3):427-442. doi: 10.1111/bjh.18407. Epub 2022 Aug 16.
Normal early erythropoiesis depends on the precise regulation of protein expression and phosphorylation modification. Dysregulation of protein levels or modification contributes to erythroid disorders. To date, the dynamics of protein phosphorylation profiling across human erythroid development is not fully understood. Here, we characterized quantitative proteomic and phosphoproteomic profiling by tandem mass-tagging technology. We systemically built phospho-expression profiling and expression clusters of 11 414 phosphopeptides for human early erythropoiesis. The standardization methods for multitier integrative analyses revealed multiple functional modules of phosphoproteins (e.g., regulation of the G2/M transition) and active phosphorylated signalling (e.g., cell cycle-related pathways). Our further analysis revealed that CDK family members were the main kinases that phosphorylate substrates in erythroid progenitors and identified that CDK9 played an important role in the proliferation of erythroid progenitors. Collectively, our phosphoproteomic profiling, integrative network analysis and functional studies define landscapes of the phosphoproteome and reveal signalling pathways that are involved in human early erythropoiesis. This study will serve as a valuable resource for further investigations of phosphatase and kinase functions in human erythropoiesis and erythroid-related diseases.
正常的早期红细胞生成依赖于蛋白质表达和磷酸化修饰的精确调控。蛋白质水平或修饰的失调会导致红细胞疾病。迄今为止，人类红细胞发育过程中蛋白质磷酸化谱的动态变化尚未完全了解。在此，我们通过串联质量标签技术对定量蛋白质组学和磷酸化蛋白质组学进行了表征。我们系统地构建了人类早期红细胞生成的11414个磷酸肽的磷酸化表达谱和表达簇。多层综合分析的标准化方法揭示了磷酸化蛋白质的多个功能模块（如G2/M期转换的调控）和活跃的磷酸化信号传导（如细胞周期相关途径）。我们的进一步分析表明，CDK家族成员是红细胞祖细胞中磷酸化底物的主要激酶，并确定CDK9在红细胞祖细胞的增殖中起重要作用。总的来说，我们的磷酸化蛋白质组学分析、综合网络分析和功能研究定义了磷酸化蛋白质组的格局，并揭示了参与人类早期红细胞生成的信号通路。这项研究将为进一步研究磷酸酶和激酶在人类红细胞生成及红细胞相关疾病中的功能提供宝贵资源。
