Analysis of the Candida albicans adhesin Als5p

白色念珠菌粘附素 Als5p 的分析

• 批准号: 6807048
• 负责人: Jason Malcolm Rauceo
• 金额: $ 2.68万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2006-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6807048
• 关键词: Candida albicans; amyloid proteins; cell adhesion; cell cell interaction; cell membrane; electron microscopy; extracellular matrix; fungal proteins; glycoproteins; high performance liquid chromatography; polymerase chain reaction; predoctoral investigator; spectrometry

DESCRIPTION (provided by applicant): Our long term objective is to understand the mechanisms and role of fungal adhesions in cellular interactions. Candida albicans infections remain a significant cause of morbidity and mortality, and are often inadequately controlled with the available antifungals, thus creating a serious health concern. We propose the following specific aims: 1. Test the idea that Candida albicans adhesin AIs5p undergoes a conformational shift from a non-aggregative to aggregative state following adherence to substrate. Conformational shifts will be monitored by CD spectroscopy. 2. Test the idea that AIs5p mediated aggregation features amyloid characteristics. CD spectroscopy, the binding of amyloid specific dyes, and electron microscopy will be tested on soluble AIs5p fragments and aggregates 3. Test the idea that the tandem repeat region of AIs5p mediates homotypic binding facilitating aggregation. Non-denaturing gel electrophoresis on soluble protein fragments and binding assays with a cell-surface expressed AIs5p variant will be performed. 4. Test the idea that the N-Terminal globular Ig-like domain of AIs5p mediates adhesion to substate. Binding assays with purified AIs5p Ig -like protein fragments will be performed. These experiments will help explain the behavior of a large group of proteins from other pathogens.

描述（由申请人提供）：我们的长期目标是了解真菌粘附在细胞相互作用中的机制和作用。 白色念珠菌的感染仍然是发病率和死亡率的重要原因，并且通常用可用的抗真菌剂控制不足，从而引起了严重的健康问题。 我们提出以下具体目的：1。测试白色念珠菌粘连蛋白AIS5P经历构象转移的想法，从遵守底物后，从非侵蚀性到总体状态的构象转移。 构象转移将通过CD光谱法监测。 2。测试AIS5P介导的聚集具有淀粉样蛋白特性的想法。 CD光谱，淀粉样蛋白特异性染料的结合和电子显微镜将在可溶性AIS5P片段上进行测试并聚集3。测试AIS5P的串联重复区域介导同型结合促进促进聚集的想法。 将在可溶性蛋白质片段上进行非换性凝胶电泳和用表达AIS5P变体的细胞表面的结合测定。 4。测试AIS5P的N末端球状Ig样结构域可介导粘附以取代的想法。 将进行纯化的AIS5P Ig -like蛋白片段的结合测定。 这些实验将有助于解释其他病原体的大量蛋白质的行为。