Comparison of Proteomic Responses

To facilitate antibiotic research, we have developed a compendium of proteomic responses that allows rapid classification of compounds into those with precedented and unprecedented modes of action. We gathered more than 100 proteomic response profiles of B. subtilis to treatment with antibiotics. To depict acute bacterial responses to sublethal doses of antibiotics, we used pulse-labeling of proteins that are newly synthesized after antibiotic challenge and analyze the proteome by 2D-PAGE. Subsequently, marker proteins were excised from non-radioactive gels and identified by mass spectrometry. Through compilation of response profiles, marker proteins are delineated as signatures, which are specific for inhibition of cellular processes like DNA, RNA, or protein biosynthesis, metabolic processes like fatty acid or folic acid biosynthesis, and attacks on cellular structures like the cytoplasmic membrane or the cell wall.

为推动抗生素研究，我们构建了一套蛋白质组响应汇编集，可快速将化合物划分为具有已知作用模式与全新作用模式的两类物质。我们收集了超过100份枯草芽孢杆菌（Bacillus subtilis）经抗生素处理后的蛋白质组响应图谱。为刻画细菌在抗生素亚致死剂量下的急性应激响应，我们对抗生素暴露后新合成的蛋白质实施脉冲标记，并通过双向聚丙烯酰胺凝胶电泳（2D-PAGE）分析蛋白质组。随后，我们从非放射性凝胶中切取标记蛋白，借助质谱分析法完成鉴定。通过整合各类响应图谱，我们将标记蛋白界定为特异性特征标志物，这些标志物可用于识别针对DNA、RNA或蛋白质生物合成等细胞过程的抑制作用，针对脂肪酸或叶酸生物合成等代谢过程的抑制作用，以及针对细胞质膜、细胞壁等细胞结构的攻击作用。