Targeted protein degradation by Salmonella under phagosome-mimicking culture conditions investigated using comparative peptidomics
Targeted protein degradation by Salmonella under phagosome-mimicking culture conditions investigated using comparative peptidomics
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DOI：
10.1074/mcp.m600282-mcp200
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发表时间：
2007-04-01
影响因子：
7
通讯作者：
Heffron, Fred
中科院分区：
文献类型：
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作者：
Manes, Nathan P.;Gustin, Jean K.;Heffron, Fred
肠道沙门氏菌的病原体是已知的，导致食物中毒和伤寒。由于抗药性分离株的出现和生物恐怖主义的威胁（例如食品供应的污染），研究这种细菌的需求越来越大。本研究采用比较肽组学方法对S.在丰富培养基或酸性、低镁和最低营养培养基中培养的肠血清型鼠伤寒沙门氏菌，所述培养基设计为大致模拟巨噬细胞吞噬体环境（已知沙门氏菌在其中存活）。通过使用异丙醇提取富集来自澄清的细胞裂解物的天然肽，并通过使用LC-MS/MS和LC-FTICRMS两者进行分析。我们鉴定并定量了来自682种蛋白质的5，163种肽，数据清楚地表明，与在丰富培养基中培养的沙门氏菌相比，在吞噬体模拟培养基中培养的细胞具有显著更高丰度的各种蛋白质降解产物，特别是来自核糖体蛋白的降解产物。来自相同培养物的沙门氏菌也使用传统的自下而上的蛋白质组学方法进行了分析，并且当肽组学和蛋白质组学数据一起分析时，初步鉴定了两种蛋白质簇，其靶向为蛋白质水解。可能的作用，有针对性的蛋白水解吞噬沙门氏菌进行了讨论。
The pathogen Salmonella enterica is known to cause both food poisoning and typhoid fever. Because of the emergence of antibiotic-resistant isolates and the threat of bioterrorism (e.g. contamination of the food supply), there is a growing need to study this bacterium. In this investigation, comparative peptidomics was used to study S. enterica serovar Typhimurium cultured in either a rich medium or in an acidic, low magnesium, and minimal nutrient medium designed to roughly mimic the macrophage phagosomal environment (within which Salmonella are known to survive). Native peptides from cleared cell lysates were enriched by using isopropanol extraction and analyzed by using both LC-MS/MS and LC-FTICRMS. We identified and quantified 5,163 peptides originating from 682 proteins, and the data clearly indicated that compared with Salmonella cultured in the rich medium, cells cultured in the phagosome-mimicking medium had dramatically higher abundances of a wide variety of protein degradation products, especially from ribosomal proteins. Salmonella from the same cultures were also analyzed using traditional, bottom-up proteomic methods, and when the peptidomics and proteomics data were analyzed together, two clusters of proteins targeted for proteolysis were tentatively identified. Possible roles of targeted proteolysis by phagocytosed Salmonella are discussed.