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首页> 外文期刊>Infection and immunity >Quantitative Profile of the Uropathogenic Escherichia coli Outer Membrane Proteome during Growth in Human Urine
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Quantitative Profile of the Uropathogenic Escherichia coli Outer Membrane Proteome during Growth in Human Urine

机译：尿液致病性大肠杆菌外膜蛋白质组在人类尿液中生长期间的定量概况

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Outer membrane proteins (OMPs) of microbial pathogens are critical components that mediate direct interactions between microbes and their surrounding environment. Consequently, the study of OMPs is integral to furthering the understanding of host-pathogen interactions and to identifying key targets for development of improved antimicrobial agents and vaccines. In this study, we used two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and tandem mass spectrometry to characterize the uropathogenic Escherichia coli (UPEC) outer membrane subproteome; 30 individual OMPs present on the bacterial surface during growth in human urine were identified. Fluorescence difference gel electrophoresis was used to identify quantitative changes in levels of UPEC strain CFT073 OMPs during growth in urine; six known receptors for iron compounds were induced in this environment, i.e., ChuA, IutA, FhuA, IroN, IreA, and Iha. A seventh putative iron compound receptor, encoded by CFT073 open reading frame (ORF) c2482, was also identified and found to be induced in urine. Further, the induction of these seven iron receptors in human urine and during defined iron limitation was verified by using quantitative real-time PCR (qPCR). An eighth iron receptor, fepA, displayed similar induction levels under these conditions as measured by qPCR but was not identified by 2D-PAGE. Addition of 10 μM FeCl2 to human urine repressed the transcription of all eight iron receptor genes. A number of fecal-commensal, intestinal pathogenic, and uropathogenic E. coli strains all displayed similar growth rates in human urine, showing that the ability to grow in urine per se is not a urovirulence trait. Thus, human urine is an iron-limiting environment and UPEC enriches its outer membrane with iron receptors to contend with this iron limitation.
机译：微生物病原体的外膜蛋白（OMP）是介导微生物与其周围环境之间直接相互作用的关键成分。因此，对OMP的研究对于增进对宿主-病原体相互作用的了解以及确定开发改良的抗菌剂和疫苗的关键目标是不可或缺的。在这项研究中，我们使用了二维聚丙烯酰胺凝胶电泳（2D-PAGE）和串联质谱法来表征尿路致病性大肠埃希氏大肠杆菌（UPEC）外膜亚蛋白质组。鉴定了在人尿中生长期间细菌表面上存在的30种OMP。荧光差异凝胶电泳用于鉴定尿液生长过程中UPEC菌株CFT073 OMPs水平的定量变化。在此环境中诱导了六个已知的铁化合物受体，即ChuA，IutA，FhuA，IroN，IreA和Iha。还确定了由CFT073开放阅读框（ORF）c2482编码的第七种假定的铁化合物受体，并在尿液中被诱导。此外，通过使用定量实时PCR（qPCR）验证了这七个铁受体在人尿中和限定的铁限量期间的诱导。通过qPCR测量，第八种铁受体 fepA 在这些条件下显示出相似的诱导水平，但未通过2D-PAGE鉴定。在人类尿液中添加10μMFeCl 2 抑制了所有八个铁受体基因的转录。许多粪便，肠道病原和尿毒症 E。大肠杆菌菌株在人尿液中均显示出相似的生长速率，表明尿液本身的生长能力并不是尿毒力的特征。因此，人类尿液是铁限制环境，UPEC的外膜富含铁受体以应对这种铁限制。

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