Proteomics of Porphyromonas gingivalis interactions with Fusobacterium nucleatum

牙龈卟啉单胞菌与具核梭杆菌相互作用的蛋白质组学

• 批准号: 8471012
• 负责人: Murray Hackett
• 金额: $ 53.97万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8471012
• 关键词: Address; Animal Model; Bacteria; Bacterial Proteins; Biochemical Pathway; Biological; Biological Models; Cell Communication; Cell Line; Cells; Chemicals; Clinical; Communities; Community Developments; Computer Simulation; Data Set; Dental; Developed Countries; Diagnostic; Disease; Epithelial Cells; Event; Fusobacterium nucleatum; Gene Expression; Gene Expression Regulation; Genes; Genetic; Gingiva; Goals; Grant; Individual; Measures; Microbe; Microbial Biofilms; Microbiology; Mining; Modeling; Modification; Molecular; NIH Program Announcements; Oral; Oral cavity; Organism; Output; Paint; Pathogenesis; Pathogenicity; Pathway interactions; Pattern; Peer Review; Periodontal Diseases; Phenotype; Phosphorylation; Porphyromonas gingivalis; Post-Translational Protein Processing; Process; Progress Reports; Proteins; Proteome; Proteomics; Relative (related person); Research; Resource Sharing; Role; Running; Science; Side; Streptococcus gordonii; Surface; System; Systems Biology; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Tooth Loss; Virulence Factors; Writing; base; comparative; data mining; experience; improved; in vivo; method development; microbial; microbial community; microorganism; microorganism interaction; mutant; novel; novel strategies; oral bacteria; oral biofilm; pathogen; protein expression; protein function; public health relevance; research study; response; transcriptomics

DESCRIPTION (provided by applicant): This project will examine global protein expression changes in oral microbial communities and the community response to model host cells. We will select as model organisms the periodontal pathogen P. gingivalis, along with F. nucleatum and S. gordonii as representative of organism commonly found in dental biofilms and capable of synergistic interactions with P. gingivalis. In addition to differential protein expression as these organisms assemble into communities, we will also determine changes in protein expression induced by contact with gingival epithelial cells. The long-term goals of the study involve improving our fundamental understanding at the molecular level of events surrounding heterotypic oral biofilm formation and the interaction of these biofilms with host cells. Specific Aim 1 defines reference proteomes of P. gingivalis, S. gordonii, and F. nucleatum individually, pairwise and all together. This will include global protein modification analysis using data mining techniques, and a chemical approach to global phosphorylation specifically. We will define protein relative abundance changes (relative to the single organism in isolation) that can be attributed to community microbial interactions. We will thus establish the proteome of the P. gingivalis-F. nucleatum-S. gordonii community, and the differential responses to community development in comparison to individual proteomes. Specific Aim 2 will define the effects of gingival epithelial cells on the P. gingivalis-F. nucleatum-S. gordonii community. This will involve running the same experiments as in Aim 1, but in the presence of epithelial cells, also with comparative posttranslational modification (PTM) analysis on the microbial side. This will define the response of the community to the presence of host cells. As in Aim 1, this includes mining the datasets globally for PTMs for all known modifications using computational approaches, and in addition using a chemically based experimental approach to measuring differential global phosphorylation. In Aim 3 we will prioritize the regulated proteins identified in Aims 1 and 2, as discovery warrants, for additional corroboration and for construction of mutants in the corresponding genes. The phenotype of the mutants in community development and responses to epithelial cells will further elucidate functional meaning and biological relevance.

描述（由申请人提供）：本项目将检查口腔微生物群落中的整体蛋白质表达变化以及对模型宿主细胞的群落反应。我们将选择牙周致病菌牙龈卟啉单胞菌作为模式生物，沿着的还有F. nucleatum和S. gordonii作为通常在牙齿生物膜中发现的生物体的代表，并且能够与牙龈卟啉单胞菌协同相互作用。除了差异蛋白质表达，因为这些生物体组装成社区，我们还将确定蛋白质表达的变化与牙龈上皮细胞接触诱导。该研究的长期目标包括提高我们在分子水平上对异型口腔生物膜形成以及这些生物膜与宿主细胞相互作用的基本理解。具体目标1定义了牙龈卟啉单胞菌、S. gordonii和F.单个、成对和一起的具核质。这将包括使用数据挖掘技术的全球蛋白质修饰分析，以及专门针对全球磷酸化的化学方法。我们将定义蛋白质相对丰度的变化（相对于孤立的单一生物体），可以归因于社区微生物的相互作用。因此，我们将建立牙龈卟啉单胞菌-F的蛋白质组。核-S。gordonii群落，以及与个体蛋白质组相比群落发展的差异反应。具体目标2将定义牙龈上皮细胞对牙龈卟啉单胞菌-F的影响。核-S。戈登社区这将涉及运行与目标1相同的实验，但在上皮细胞存在下，还在微生物方面进行比较翻译后修饰（PTM）分析。这将定义群落对宿主细胞存在的反应。如在目标1中，这包括使用计算方法全局地挖掘PTM的数据集以用于所有已知的修饰，并且另外使用基于化学的实验方法来测量差异全局磷酸化。在目标3中，我们将优先考虑目标1和2中鉴定的受调控蛋白，作为发现证明，用于额外的确证和在相应基因中构建突变体。突变体在群落发育和对上皮细胞的反应中的表型将进一步阐明功能意义和生物学相关性。