用于鉴定沙门氏菌中短链脂肪酸修饰蛋白的化学蛋白质组学方法。
Chemical proteomics for identifying short-chain fatty acid modified proteins in Salmonella.
机构信息
Department of Immunology and Microbiology, Scripps Research, La Jolla, CA, United States.
Duchossois Family Institute, The University of Chicago, Chicago, IL, United States.
出版信息
Methods Enzymol. 2022;664:135-150. doi: 10.1016/bs.mie.2021.11.012. Epub 2021 Dec 21.
Microbiota-metabolized small molecules play important roles to regulate host immunity and pathogen virulence. Specifically, microbiota generates millimolar concentration of short-chain fatty acid (SCFA) that can directly inhibit Salmonella virulence. Here, we describe chemical proteomic methods to identify SCFA-modified proteins in Salmonella using free fatty acids as well as their salicylic acid derivatives. In addition, we include CRISPR-Cas9 gene editing protocols for epitope-tagging of specific proteins to validate SCFA-modification in Salmonella. These protocols should facilitate the discovery and functional analysis of SCFA-modified proteins in Salmonella microbiology and pathogenesis.
微生物群代谢的小分子在调节宿主免疫和病原体毒力方面发挥着重要作用。具体来说，微生物群产生毫摩尔浓度的短链脂肪酸 (SCFA)，可以直接抑制沙门氏菌的毒力。在这里，我们描述了使用游离脂肪酸及其水杨酸衍生物来鉴定沙门氏菌中 SCFA 修饰蛋白的化学蛋白质组学方法。此外，我们还包括 CRISPR-Cas9 基因编辑方案，用于对特定蛋白质进行表位标记，以验证沙门氏菌中 SCFA 的修饰。这些方案应该有助于发现和功能分析沙门氏菌微生物学和发病机制中的 SCFA 修饰蛋白。
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