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）是人类中最常见的细菌感染之一，到目前为止，UTI最常见的唯一病因是大肠杆菌。膀胱感染的一个关键方面是细菌侵袭膀胱上皮。最近，我们发现侵袭人膀胱上皮和随后的细菌转胞细胞增多症是由称为Caveolae的不同血浆细胞实体进行了批判性介导的。发现1型纤维脑病发育性大肠杆菌通过结合Cavolae的成分侵入膀胱细胞。大肠杆菌通过Caveolae入侵被包裹在小膜中，并经过转移而不会丧失生存能力。这些观察结果表明小窝是进入和感染的重要门户。目前对膀胱细胞内的洞穴结构以及如何被病原体进行进入的洞穴结构很少。也不知道是什么使caveolol蛋白中的大肠杆菌与进入后的溶酶体融合在一起。在细菌激活之前和之后，确定膀胱上皮细胞中小窝的蛋白质组成可能会获得有关这些问题的有价值的信息。在这里，我们建议：（i）使用蛋白质组采矿方法来鉴定与大肠杆菌相互作用之前和之后，在人膀胱上皮细胞中发现的小窝的蛋白质组成。我们的重点将放在作用为（a），受体（b），信号分子和（c），细胞骨架重排和运输的介体的分子上。 （ii），通过特异性抑制基因表达并检查其对E.Coli的结合，侵袭和/或细胞内运输活性的影响来推断选定的洞穴蛋白的功能。这些提出的研究涉及探索技术（蛋白质组学）的利用，这些研究将补充目前正在进行的NIH资助研究。