全球磷酸化蛋白质组学谱揭示了B细胞非霍奇金淋巴瘤的鲜明特征 Am J Pathol. 2014,84(5):1331-42.
在造血系统恶性肿瘤的发病机制中，信号通路的反常受蛋白磷酸化的调控；某种程度上讲，磷酸化的失控可能与B细胞非霍奇金淋巴瘤（B-NHL）的发病机制有关。为了鉴定磷酸化在B-NHLs发病过程中的重要性，我们对起源于3类B-NHL （包括伯基特淋巴瘤、滤泡性淋巴瘤和套细胞淋巴瘤）的11种细胞进行了质谱分析、label-free和半定量磷酸化蛋白质组学分析。我们对6579独特的磷酸化多肽和相应的1701磷酸化蛋白进行了鉴定和量化。在ProteomeXchange数据库中通过识别编码PXD000658可以获得这些数据，每一个淋巴瘤亚型高度显示了不同的磷酸化特征。有趣的是，在生发中心起源的B-NHL细胞系中，蛋白的磷酸化参与了B细胞受体信号通路。这些蛋白中，与磷酸化蛋白相关的PAG1被鉴定为与伯基特淋巴瘤和滤泡性淋巴瘤关系最为密切的磷酸化酪氨酸多肽。下调PAG1会导致B细胞受体信号通路中酪氨酸磷酸化过程受阻，且能够明显的增加生发中心起源的B-NHLs细胞增殖和对抗原刺激的反应。这些数据对磷酸化蛋白在人类淋巴瘤发生过程中所起的作用提供了详细的注解。总的来说，我们的研究揭示了磷酸化蛋白质组学在信号通路中的特征，并对了解B细胞淋巴的发病机制提出了新的认识。

Deregulation of signaling pathways controlled by protein phosphorylation underlies the pathogenesis of hematological malignancies; however, the extent to which deregulated phosphorylation may be involved in B- cell non-Hodgkin lymphoma (B-NHL ) pathogenesis is la rgely unknown . To identify phosphorylation events important in B-NHLs, we performed mass spectrometry-based,label-free, semiquantitative phosphoproteomic profiling of 11 cell lines d erived from three B-NHL categories: Burkitt lymphoma, follicular lymphoma, and mantle-cell lymphoma. In all, 6579 unique phosphopeptides, corresponding to 1701 unique phosphorylated proteins, were identified and quantified. The data are available via ProteomeXchange with identifier PXD000658 . Hierarchical clustering highlighted distinct phosphoproteomic signatures associated with each lymphoma subtype. Interestingly, germinal center-derived B-NHL cell lines were characterized by phosphorylation of proteins involved in the B-cell receptor signaling. Of these proteins, phosphoprotein associated with glycosphingolipid - enriched microdomains 1 ( PAG1) was idetified with the most phosphorylated tyrosine peptides in Burkitt lymphoma and follicular lymphoma. PAG1 knockdown resulted in perturbation of the tyrosine phosphosignature of B -cell receptor signaling components. Significantly, PAG1 knockdown increased cell proliferation and response to antigen stimulation of these germinal center-derived B -NHLs. These data provide a detailed annotation of phosphorylated proteins in human lymphoid cancer . Over all , our study revealed the utility of unbiased phosphoproteome interrogation in characterizing signaling networks that may provide insights into pathogenesis mechanisms in B - cell lymphomas .