Chen Xiao-Bo, Liao Dong-Jiang, Li Shi-Yue, He Ying, Chi Feng
Guangzhou Institute of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical College, Guangzhou 510120, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2009 Nov;32(11):842-6.
To screen the differential expressed proteins of peripheral blood mononuclear cells (PBMC) in patients with pulmonary tuberculosis by comparative proteomics.
PBMCs from 10 patients with tuberculosis and 10 healthy volunteers were collected. The total proteins were extracted and separated by two-dimensional gel electrophoresis (2-DE). The sites of differential expression were analyzed using ImageMaster 2D Elite 5.0 image analysis software, and the peptide mass finger-printing (PMF) identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The biological information of these proteins was searched in the database and confirmed by Western blot.
Well reproducible 2-DE maps of PBMC from the patients with tuberculosis and normal controls were established. Forteen differential protein spots were found. Twelve proteins were analyzed and identified. Pleckstrin and Ras suppressor protein1 were decreased in patients with tuberculosis while the other 10 proteins were increased. The decreased expression of Pleckstrin in tuberculosis was confirmed by Western blot.
Analysis of differential protein expression in PBMC from patients with pulmonary tuberculosis by comparative proteomics identified 12 proteins. The results suggest that Pleckstrin and Coronin-1A may have an important role in the pathogenesis of pulmonary tuberculosis through regulating the phagocytosis of mononuclear macrophages.
通过比较蛋白质组学筛选肺结核患者外周血单个核细胞(PBMC)中的差异表达蛋白。
收集10例肺结核患者和10名健康志愿者的PBMC。提取总蛋白并通过二维凝胶电泳(2-DE)进行分离。使用ImageMaster 2D Elite 5.0图像分析软件分析差异表达位点,并通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)鉴定肽质量指纹图谱(PMF)。在数据库中搜索这些蛋白质的生物学信息,并通过蛋白质印迹法进行确认。
建立了肺结核患者和正常对照者PBMC的可重复的2-DE图谱。发现14个差异蛋白点。分析并鉴定了12种蛋白质。肺结核患者中Pleckstrin和Ras抑制蛋白1减少,而其他10种蛋白质增加。蛋白质印迹法证实了肺结核中Pleckstrin的表达降低。
通过比较蛋白质组学分析肺结核患者PBMC中的差异蛋白表达,鉴定出12种蛋白质。结果表明,Pleckstrin和冠蛋白-1A可能通过调节单核巨噬细胞的吞噬作用在肺结核发病机制中起重要作用。