Pförtner Henrike, Depke Maren, Surmann Kristin, Schmidt Frank, Völker Uwe
Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, Germany.
Methods Mol Biol. 2018;1841:207-228. doi: 10.1007/978-1-4939-8695-8_15.
Proteome profiling of bacteria internalized by host cells is still a challenging task, due to low amounts of bacterial proteins in host-pathogen settings and the high amounts of contaminating host proteins. Here, we describe a workflow for the enrichment of intracellular bacteria by fluorescence activated cell sorting which in combination with highly sensitive LC-MS/MS allows monitoring of about 1200 proteins from 2 to 4 × 10 internalized bacterial cells as starting material.
由于在宿主 - 病原体环境中细菌蛋白含量低且宿主污染蛋白含量高，对宿主细胞内化细菌进行蛋白质组分析仍然是一项具有挑战性的任务。在此，我们描述了一种通过荧光激活细胞分选富集细胞内细菌的工作流程，该流程与高灵敏度液相色谱 - 串联质谱联用，能够以2至4×10个内化细菌细胞作为起始材料监测约1200种蛋白质。
