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目的 试图运用蛋白组学的策略研究系统性红斑狼疮 (SLE)相关的干扰素诱导蛋白IFIT1的蛋白质相互作用 ,以揭示其可能的功能。方法 应用荧光实时定量PCR进一步验证SLE患者外周血白细胞转录水平IFIT1基因的表达 ;应用分子克隆技术在大肠杆菌克隆并表达谷胱甘肽S转移酶 (GST) IFIT1融合蛋白 ;利用GST表达标签 ,以GST IFIT1为饵蛋白捕获SLE外周血白细胞蛋白裂解物中与IFIT1存在相互作用的蛋白质 ,MALDI TOF MS肽指纹图谱进行蛋白质鉴定。结果 荧光实时定量PCR证实IFIT1基因确在SLE(4 0例 )较正常对照 (2 9例 )表达上调 (P <0 0 0 1)。克隆表达了GST IFIT1融合蛋白 ,并捕获到一个与IFIT1存在相互作用的蛋白质 ,经肽指纹图谱初步鉴定为Rho/Rac鸟嘌呤核苷交换因子。结论 IFIT1是SLE新的相关基因。运用蛋白组学的策略研究IFIT1蛋白质水平的相互作用显示 ,IFIT1与Rho/Rac鸟嘌呤核苷交换因子存在相互作用 ,IFIT1可能通过该途径影响Rho蛋白的活化 ,参与SLE的发病。
OBJECTIVE: To study the protein interaction of systemic lupus erythematosus (SLE) -related interferon-inducible protein IFIT1 using proteomic strategies to reveal its possible function. Methods Real-time quantitative PCR was used to further verify the expression of IFIT1 gene in peripheral blood leukocytes of SLE patients. Clone of glutathione S-transferase (GST) IFIT1 fusion protein was cloned by E. coli using molecular cloning technology. GST IFIT1 captures proteins interacting with IFIT1 in SLE peripheral blood leukocyte lysates as bait proteins and MALDI TOF MS peptide fingerprinting for protein identification. Results Fluorescence real-time quantitative PCR confirmed that IFIT1 gene was up-regulated in SLE (29 cases) compared with normal controls (29 cases) (P <0.01). The GST IFIT1 fusion protein was cloned and cloned, and a protein interacting with IFIT1 was captured and preliminarily identified as Rho / Rac guanosine exchange factor by peptide fingerprinting. Conclusion IFIT1 is a novel related gene of SLE. Using proteomics strategy to study the interaction of IFIT1 protein levels shows that IFIT1 interacts with Rho / Rac guanosine exchange factor, and IFIT1 may affect the activation of Rho protein through this pathway and participate in the pathogenesis of SLE.