Department of Nephrology, Shenzhen People's Hospital, Second Clinical Medical College, Jinan University, Shenzhen, China.
PLoS One. 2012;7(12):e53129. doi: 10.1371/journal.pone.0053129. Epub 2012 Dec 28.
Protein phosphorylation is a complex regulatory event that is involved in the signaling networks that affect virtually every cellular process. The protein phosphorylation may be a novel source for discovering biomarkers and drug targets. However, a systematic analysis of the phosphoproteome in patients with SLE has not been performed. To clarify the pathogenesis of systemic lupus erythematosus (SLE), we compared phosphoprotein expression in PBMCs from SLE patients and normal subjects using proteomics analyses. Phosphopeptides were enriched using TiO₂ from PBMCs isolated from 15 SLE patients and 15 healthy subjects and then analyzed by automated LC-MS/MS analysis. Phosphorylation sites were identified and quantitated by MASCOT and MaxQuant. A total of 1035 phosphorylation sites corresponding to 618 NCBI-annotated genes were identified in SLE patients compared with normal subjects. Differentially expressed proteins, peptides and phosphorylation sites were then subjected to bioinformatics analyses. Gene ontology(GO) and pathway analyses showed that nucleic acid metabolism, cellular component organization, transport and multicellular organismal development pathways made up the largest proportions of the differentially expressed genes. Pathway analyses showed that the mitogen-activated protein kinase (MAPK) signaling pathway and actin cytoskeleton regulators made up the largest proportions of the metabolic pathways. Network analysis showed that rous sarcoma oncogene (SRC), v-rel reticuloendotheliosis viral oncogene homolog A (RELA), histone deacetylase (HDA1C) and protein kinase C, delta (PRKCD) play important roles in the stability of the network. These data suggest that aberrant protein phosphorylation may contribute to SLE pathogenesis.
蛋白质磷酸化是一种复杂的调节事件，涉及影响几乎所有细胞过程的信号转导网络。蛋白质磷酸化可能是发现生物标志物和药物靶点的新来源。然而，尚未对 SLE 患者的磷酸蛋白质组进行系统分析。为了阐明系统性红斑狼疮（SLE）的发病机制，我们使用蛋白质组学分析比较了 SLE 患者和正常对照者 PBMC 中的磷酸蛋白表达。从 15 例 SLE 患者和 15 例健康对照者分离的 PBMC 中使用 TiO₂ 富集磷酸肽，然后通过自动 LC-MS/MS 分析进行分析。通过 MASCOT 和 MaxQuant 鉴定和定量磷酸化位点。与正常对照者相比，在 SLE 患者中鉴定出 1035 个对应于 618 个 NCBI 注释基因的磷酸化位点。然后对差异表达蛋白、肽和磷酸化位点进行生物信息学分析。GO 和途径分析显示，核酸代谢、细胞成分组织、运输和多细胞生物发育途径构成了差异表达基因的最大比例。途径分析显示，丝裂原活化蛋白激酶（MAPK）信号通路和肌动蛋白细胞骨架调节剂构成了代谢途径的最大比例。网络分析显示，劳斯肉瘤癌基因（SRC）、v-rel 网状内皮增生病毒癌基因同源物 A（RELA）、组蛋白脱乙酰酶（HDA1C）和蛋白激酶 C，δ（PRKCD）在网络稳定性中发挥重要作用。这些数据表明，异常的蛋白质磷酸化可能导致 SLE 的发病机制。
