Proteome Mining in Caveolae of the Bladder Epithelium

膀胱上皮细胞小窝的蛋白质组挖掘

• 批准号: 6779266
• 负责人: Soman N Abraham
• 金额: $ 15.4万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6779266
• 关键词: Escherichia coli; Escherichia coli infections; bacteria infection mechanism; bacterial cytopathogenic effect; bioinformatics; caveolas; cell component structure /function; computational biology; cytoskeleton; gene expression; high throughput technology; host organism interaction; intracellular transport; mass spectrometry; protein quantitation /detection; protein structure; protein structure function; proteomics; two dimensional gel electrophoresis; urinary bladder epithelium; urinary tract infection

DESCRIPTION (provided by applicant): Urinary Tract Infections (UTIs) are one of the most common bacterial infections in man, and by far the single most common causative agent of UTIs is Escherichia coli. A critical aspect of bladder infection is bacterial invasion of the bladder epithelium. Recently, we discovered that invasion of human bladder epithelium and subsequent bacterial transcytosis was critically mediated by distinct plasmalemmal cellular entities called caveolae. Type 1 fimbriated uropathogenic E.coli were discovered to invade bladder cells by binding components of cavolae. E.coli invading via caveolae are encased in caveolar membranes and transcytozed without loss of viability. These observations point to caveolae as important portals for entry and infection. Very little is currently known of the caveolar structure within bladder cells, and how it is co-opted by pathogens for entry. It is also not known what keeps E.coli encased in caveolar proteins from fusing with lysosomes following entry. Determining the protein composition of caveolae in bladder epithelial cells before and after bacterial activation could derive valuable information regarding these issues. Here we propose to: (i) Use proteome mining approaches to identify protein composition of caveolae found in human bladder epithelial cells before and after interaction with E.coli. Our focus will be on molecules functioning as (a), receptors (b), signaling molecules and (c), mediators of cytoskeletal rearrangement and trafficking. (ii), Deduce the functions of selected caveolar proteins by specifically suppressing gene expression and examining its effects on the binding, invasion and/or intracellular trafficking activities of the E.coli. These proposed studies involve the utilization of exploratory technology (proteomics), which will complement other NIH funded research currently being undertaken.

描述（由申请人提供）：尿路感染（UTI）是人类最常见的细菌感染之一，迄今为止，UTI 最常见的单一病原体是大肠杆菌。膀胱感染的一个重要方面是细菌侵入膀胱上皮。最近，我们发现人类膀胱上皮的侵袭和随后的细菌转胞吞作用关键是由称为小窝的不同质膜细胞实体介导的。 1 型伞状尿路致病性大肠杆菌被发现通过结合卡沃拉的成分来侵入膀胱细胞。通过细胞膜穴入侵的大肠杆菌被包裹在细胞膜穴膜中并进行转胞吞，而不会丧失活力。这些观察结果表明，小窝是进入和感染的重要门户。目前，人们对膀胱细胞内的小凹结构以及病原体如何选择其进入的了解甚少。目前还不清楚是什么阻止了包裹在小凹蛋白中的大肠杆菌在进入溶酶体后不与溶酶体融合。确定细菌激活前后膀胱上皮细胞中小窝的蛋白质组成可以得出有关这些问题的有价值的信息。在这里，我们建议：（i）使用蛋白质组挖掘方法来鉴定在与大肠杆菌相互作用之前和之后在人类膀胱上皮细胞中发现的小窝的蛋白质组成。我们的重点将是发挥（a）受体（b）信号分子和（c）细胞骨架重排和运输介体作用的分子。 (ii)，通过特异性抑制基因表达并检查其对大肠杆菌的结合、入侵和/或细胞内运输活动的影响来推断选定的小凹蛋白的功能。这些拟议的研究涉及探索性技术（蛋白质组学）的利用，这将补充目前正在进行的其他 NIH 资助的研究。