A proteomics approach to Candida albicans biofilm

白色念珠菌生物膜的蛋白质组学方法

• 批准号: 6782705
• 负责人: Welda LaJean CHAFFIN
• 金额: $ 3.38万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6782705
• 关键词: Candida albicans; Europe; biofilm; biotechnology; cell wall; gene expression; mass spectrometry; matrix assisted laser desorption ionization; polymerase chain reaction; protein structure function; proteomics; saliva

DESCRIPTION (provided by applicant): This research will be done primarily in Spain at Universidad de Complutense Madrid in collaboration with Dr. Cesar Nombela as an extension of NIH grant R01 DE14029. The hypothesis of the parent grant and this FIRCA application is that alterations in gene expression accompany the change of Candida albicans from a free living planktonic cell to a sessile member of a biofilm. The parent grant focuses on expression profiling (microarrays) in mono- and mixed species biofilm and in organisms recovered from the oral cavity (Aims 1 and 2) and analysis of selected regulated genes through generation of deficient mutant strains (Aim 3). This FIRCA project is a proteomic project to examine protein expression and identify differences between planktonic and sessile organisms. Nombela and colleagues have established the feasibility of a proteomics approach with studies of proteins in cell wall subfractions of C. albicans yeast cells and germ tubes and have used mass spectrophotometric (MS) techniques (MALDI-TOF) for identification of cell wall proteins from Saccharomyces cerevisiae and C. albicans cytoplasm Cell wall and cytoplasmic extracts will be examined and, time permitting other subcellular fractions may be examined if indicated by expression analysis derived from the parent grant. In Aim 1, the profiles of extracts from biofilm organisms will be compared to that of yeast cells and germ tubes. This complements the first aim of the parent grant. In Aim 2, the examination will be extended to extracts from selected mutant strains. This complements the third aim of the parent grant in which the mutant strains are constructed and examined for biofilm formation and global gene expression. Proteins will be separated by 2D-PAGE and profiles compared to identify qualitative and quantitative differences in protein abundance. Proteins that differ between planktonic and biofilm organisms or wild type and mutant strain will be identified by mass spectrometric methods with priority given to differences in Aim 1 profiles.

描述（由申请人提供）： 这项研究将主要在西班牙马德里康普顿斯大学与 Cesar Nombela 博士合作进行，作为 NIH 拨款 R01 DE14029 的延伸。父母资助和本 FIRCA 申请的假设是，基因表达的改变伴随着白色念珠菌从自由生活的浮游细胞到生物膜的固着成员的变化。母基金的重点是单一和混合物种生物膜以及从口腔中回收的生物体中的表达谱（微阵列）（目标1和2），以及通过生成缺陷突变株来分析选定的调节基因（目标3）。该 FIRCA 项目是一个蛋白质组学项目，旨在检查蛋白质表达并识别浮游生物和无柄生物之间的差异。 Nombela 及其同事通过研究白色念珠菌酵母细胞和胚芽管的细胞壁亚级分中的蛋白质，确定了蛋白质组学方法的可行性，并使用质谱 (MS) 技术 (MALDI-TOF) 鉴定来自酿酒酵母和白色念珠菌细胞质的细胞壁蛋白质。将对细胞壁和细胞质提取物进行检查 并且，如果来自亲本授权的表达分析表明，可以检查时间允许的其他亚细胞部分。在目标 1 中，生物膜生物提取物的概况将与酵母细胞和胚芽管的概况进行比较。这补充了家长补助金的第一个目标。在目标 2 中，检查将扩展到选定突变菌株的提取物。这补充了亲本资助的第三个目标，即构建突变菌株并检查生物膜形成和全局基因表达。将通过 2D-PAGE 分离蛋白质并进行比较，以确定蛋白质丰度的定性和定量差异。将通过质谱方法鉴定浮游生物和生物膜生物或野生型和突变株之间存在差异的蛋白质，并优先考虑 Aim 1 谱中的差异。