Chemical proteomics for identifying short-chain fatty acid modified proteins in Salmonella.

Chemical proteomics for identifying short-chain fatty acid modified proteins in Salmonella.
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DOI:
10.1016/bs.mie.2021.11.012
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发表时间:
2022
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中科院分区:
生物学4区
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微生物区系代谢的小分子在调节宿主免疫和病原菌毒力方面起着重要作用。具体地说,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修饰的蛋白质的化学蛋白质组学方法。此外,我们还包括CRISPR-Cas9基因编辑协议,用于特定蛋白质的表位标记,以验证沙门氏菌中SCFA的修饰。这些方案将有助于沙门氏菌微生物学和发病机制中SCFA修饰蛋白的发现和功能分析。
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.
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