尿路致病性大肠杆菌在人尿和J82膀胱细胞中的蛋白质组学研究

IF 4 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Proteomes Pub Date : 2022-05-05 DOI:10.3390/proteomes10020015
S. Andersen, A. Nawrocki, A. E. Johansen, Ana Herrero-Fresno, Vanesa García Menéndez, J. Møller-Jensen, J. E. Olsen
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引用次数: 3

摘要

尿路致病性大肠杆菌(UPEC)是尿路感染(UTI)最常见的原因。UPEC通常存在于肠道中,在UTI形成过程中,它们经历代谢适应,首先是尿液,然后是组织侵入膀胱细胞内部。为了了解这些适应性,我们使用定量蛋白质组学分析来表征UPEC菌株UTI89在人类尿液中生长和在J82膀胱细胞内生长时的蛋白质表达。为了便于在膀胱细胞中检测UPEC蛋白而不是过量的真核蛋白,我们开发了一种方法,将生长在MOPS和尿液中的UTI89蛋白刺入以增强细菌蛋白的检测。检测到2000多种大肠杆菌蛋白。在尿液生长过程中,与铁获取和几种氨基酸摄取和生物合成系统相关的蛋白质,最显著的是精氨酸代谢,显著上调。在J82细胞的生长过程中,与铁摄取和精氨酸代谢相关的蛋白质以及与硫化合物转换相关的蛋白质也同样上调。在这种环境下,核糖体蛋白相对于MOPS的生长被下调。上调的蛋白质和被报道为感染所必需的蛋白质之间没有直接的联系,这表明生长过程中的上调并不意味着蛋白质在某种条件下是生长所必需的。
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Proteomes of Uropathogenic Escherichia coli Growing in Human Urine and in J82 Urinary Bladder Cells
Uropathogenic Escherichia coli (UPEC) are the most common cause of urinary tract infection (UTI). UPEC normally reside in the intestine, and during establishment of UTI, they undergo metabolic adaptations, first to urine and then upon tissue invasion to the bladder cell interior. To understand these adaptations, we used quantitative proteomic profiling to characterize protein expression of the UPEC strain UTI89 growing in human urine and when inside J82 bladder cells. In order to facilitate detection of UPEC proteins over the excess amount of eukaryotic proteins in bladder cells, we developed a method where proteins from UTI89 grown in MOPS and urine was spiked-in to enhance detection of bacterial proteins. More than 2000 E. coli proteins were detected. During growth in urine, proteins associated with iron acquisition and several amino acid uptake and biosynthesis systems, most prominently arginine metabolism, were significantly upregulated. During growth in J82 cells, proteins related to iron uptake and arginine metabolisms were likewise upregulated together with proteins involved in sulfur compound turnover. Ribosomal proteins were downregulated relative to growth in MOPS in this environment. There was no direct correlation between upregulated proteins and proteins reported to be essential for infections, showing that upregulation during growth does not signify that the proteins are essential for growth under a condition.
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来源期刊
Proteomes
Proteomes Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.50
自引率
3.00%
发文量
37
审稿时长
11 weeks
期刊介绍: Proteomes (ISSN 2227-7382) is an open access, peer reviewed journal on all aspects of proteome science. Proteomes covers the multi-disciplinary topics of structural and functional biology, protein chemistry, cell biology, methodology used for protein analysis, including mass spectrometry, protein arrays, bioinformatics, HTS assays, etc. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. Therefore, there is no restriction on the length of papers. Scope: -whole proteome analysis of any organism -disease/pharmaceutical studies -comparative proteomics -protein-ligand/protein interactions -structure/functional proteomics -gene expression -methodology -bioinformatics -applications of proteomics
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