School of Biosciences, University of Birmingham , Edgbaston, Birmingham, B15 2TT, United Kingdom.
Anal Chem. 2014 Nov 4;86(21):10504-10. doi: 10.1021/ac503349d. Epub 2014 Oct 23.
Top-down identification of proteins by liquid extraction surface analysis (LESA) mass spectrometry has previously been reported for tissue sections and dried blood spot samples. Here, we present a modified "contact" LESA method for top-down analysis of proteins directly from living bacterial colonies grown in Petri dishes, without any sample pretreatment. It was possible to identify a number of proteins by use of collision-induced dissociation tandem mass spectrometry followed by searches of the data against an E. coli protein database. The proteins identified suggest that the method may provide insight into the bacterial response to environmental conditions. Moreover, the results show that the "contact" LESA approach results in a smaller sampling area than typical LESA, which may have implications for spatial profiling.
此前，已有研究报道过采用液提取表面分析（LESA）质谱法对组织切片和干血斑样本进行自上而下的蛋白质鉴定。在此，我们提出了一种改良的“接触式”LESA 方法，可直接对培养在培养皿中的活体细菌菌落进行自上而下的蛋白质分析，无需任何样本预处理。通过使用碰撞诱导解离串联质谱法对其进行分析，并将数据与大肠杆菌蛋白质数据库进行搜索，我们得以鉴定出一些蛋白质。鉴定出的蛋白质表明，该方法可能有助于深入了解细菌对环境条件的反应。此外，结果表明，“接触式”LESA 方法的采样面积比典型的 LESA 小，这可能对空间分析有影响。
