Kuhn Karsten, Baumann Christian, Tommassen Jan, Prinz Thorsten
Proteome Sciences R&D, Frankfurt am Main, Germany.
Methods Mol Biol. 2012;799:127-41. doi: 10.1007/978-1-61779-346-2_8.
In addition to standard gel-based proteomic approaches, gel-free approaches using isobaric label reagents, such as Tandem Mass Tags (TMT), provide a straightforward method for studying adaptations in microbial proteomes to changing environmental conditions. This approach does not have the known difficulties of 2-D gel electrophoresis with proteins of extreme biochemical properties. The workflow described here was designed to study adaptive responses in bacteria and has been applied to study the response of meningococci to iron limitation. The supplemental use of western blotting allows the confirmation of certain changes in protein abundance identified within the TMT study.
除了基于凝胶的标准蛋白质组学方法外，使用等压标记试剂（如同位素标记串联质谱标签，TMT）的无凝胶方法为研究微生物蛋白质组对不断变化的环境条件的适应性提供了一种直接的方法。这种方法不存在二维凝胶电泳在处理具有极端生化特性蛋白质时所面临的已知困难。这里描述的工作流程旨在研究细菌中的适应性反应，并已应用于研究脑膜炎球菌对铁限制的反应。蛋白质免疫印迹法的补充使用能够确认在TMT研究中所识别出的蛋白质丰度的某些变化。
