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: 6906516
• 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 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6906516
• 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与塞萨尔·诺姆贝拉（Cesar Nombela）博士合作作为NIH Grant R01 DE14029的扩展。父母赠款和这种FIRCA应用的假设是，基因表达的改变伴随着白色念珠菌从自由活的浮游细胞变为生物膜的无柄成员。父母的赠款专注于单物种和混合物种生物膜中的表达分析（微阵列）以及从口腔中回收的生物（目标1和2）以及通过产生缺陷突变菌株对所选调节基因的分析（AIM 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确定浮游生物和无柄生物之间的差异。 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并且，如果通过父母赠款得出的表达分析表示，则可以检查允许其他亚细胞分数的时间。在AIM 1中，将比较来自生物膜生物体的提取物与酵母细胞和细菌管的提取物。这补充了父母赠款的第一个目标。在AIM 2中，检查将扩展到从选定的突变菌株中提取。这补充了父母赠款的第三个目标，在该目标中，突变菌株被构造并检查生物膜形成和全球基因表达。与确定蛋白质丰度的质量和定量差异相比，蛋白质将通过2D-PAGE和曲线分离。浮游生物和生物膜生物或野生型和突变菌株之间不同的蛋白质将通过质谱方法鉴定，并且优先考虑AIM 1曲线的差异。

项目成果

Cell surface shaving of Candida albicans biofilms, hyphae, and yeast form cells.

白色念珠菌生物膜、菌丝和酵母形成细胞的细胞表面削片。

• DOI: 10.1002/pmic.201100588
• 发表时间: 2012
• 期刊: Proteomics
• 影响因子: 3.4
• 作者: Vialas,Vital;Perumal,Palani;Gutierrez,Dolores;Ximenez-Embun,Pilar;Nombela,Cesar;Gil,Concha;Chaffin,WLaJean
• 通讯作者: Chaffin,WLaJean