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目的以双向电泳结合多维色谱串联质谱策略,分析阿替洛尔对映异构体[(R)/(S)-ATN]作用于血管平滑肌细胞(vascular smooth muscle cells,VSMCs)后的差异表达蛋白质。方法以(R)/(S)-ATN分别作用于VSMCs,提取总蛋白质,双向电泳分离差异表达蛋白点,二维纳流液相色谱芯片(HPLC-CHIP,包含一支纳升级Zorbax 300SB-C18富集柱和一支纳升级Zorbax 300SB-C18分析柱)富集、分离差异表达蛋白酶解后的肽混合物,线性离子阱质谱进行在线分析,Spectrum Mill MS Proteomics Workbench自动分析MS和MS/MS数据,通过UniProt KB/SWISS-PORT数据库进行搜索。结果经比较、筛选,(S)-ATN干预VSMCs后的蛋白质组中有6个蛋白质点表达上调,分别是CrK相关底物蛋白、接头蛋白-3复合物b-1亚基、未定性蛋白C3orf19、线粒体苹果酸脱氢酶前体1、磷酸甘油醛脱氢酶、内皮细胞清道夫受体2前体。结论 (R)/(S)-ATN可引起广泛的蛋白质表达改变,除未定性蛋白C3orf19以外,其余5种蛋白质可能与细胞信号传导与跨膜转运、细胞能量代谢以及介导乙酰化低密度脂蛋白修饰与降解过程有关。
OBJECTIVE: To study the effect of atenolol enantiomer [(R) / (S) -ATN] on the expression of differentially expressed proteins in vascular smooth muscle cells (VSMCs) by two-dimensional electrophoresis combined with multi-dimensional tandem mass spectrometry . Methods Total RNA was extracted from VSMCs with (R) / (S) -ATN. The differentially expressed protein spots were separated by two-dimensional electrophoresis. Two-dimensional nanoemulsion HPLC chips (HPLC-CHIP including a Zorbax 300SB-C18 Enriched columns and a upgraded Zorbax 300SB-C18 analytical column), separation of differentially expressed proteolysis peptide mixtures, and linear ion trap mass spectrometry for on-line analysis. Spectrum Mill MS Proteomics Workbench automatically analyzes MS and MS / MS data, Search through the UniProt KB / SWISS-PORT database. RESULTS: Six protein spots were up-regulated in the proteome after VSMCs were screened by (S) -ATN, which were CrK-related substrate protein, adapter protein-3 complex b-1 subunit and untreated protein C3orf19 , Mitochondrial malate dehydrogenase precursor 1, glyceraldehyde phosphate dehydrogenase, endothelial cell scavenger receptor 2 precursor. Conclusion (R) / (S) -ATN can cause a wide range of protein expression changes, in addition to the unidentified protein C3orf19, the remaining five proteins may be involved in cell signaling and transmembrane transport, cellular energy metabolism, and mediated acetylated low-density lipids Protein modification and degradation process.