Department of Microbial Proteomics, University of Greifswald, Institute of Microbiology, Greifswald, Germany.
University of Basel, Biozentrum, Focal Area Infection Biology, Basel, Switzerland.
Methods Mol Biol. 2023;2601:335-348. doi: 10.1007/978-1-0716-2855-3_18.
Absolute protein quantification is an essential tool for system biology approaches and elucidation of stoichiometry of multi-protein complexes. In this updated chapter, a universal protocol for gel-free absolute protein quantification in bacterial systems is described, which provides adapted methods for cytosolic and membrane proteins. This protocol can be used for sample preparation prior to miscellaneous mass spectrometry-based quantification workflows like AQUA, Hi3, and emPAI. In addition, a focus has been set to the specific challenges in antibiotic stress research.
绝对蛋白质定量是系统生物学方法和阐明多蛋白复合物计量比的重要工具。在这一更新的章节中，描述了一种用于细菌系统的无胶绝对蛋白质定量的通用方案，该方案为细胞质和膜蛋白提供了适配的方法。该方案可用于各种基于质谱的定量工作流程（如 AQUA、Hi3 和 emPAI）之前的样品制备。此外，本研究还特别关注抗生素应激研究中的具体挑战。
