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: 7779327
• 负责人: Murray Hackett
• 金额: $ 50.16万
• 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 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7779327
• 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 Regulation; Gene Proteins; 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. PUBLIC HEALTH RELEVANCE: Periodontal, or gum, diseases are the major cause of tooth loss in developed countries and result from the concerted activities of groups of microorganisms. These organisms adjust expression of their proteins as they assemble into communities and interact with host cells. We will define these protein expression changes and assess their importance for contribution to the disease process. Gaining an improved understanding of protein changes is a basic requirement for improved therapeutic and diagnostic strategies.

描述（由申请人提供）：该项目将检查口腔微生物群落的整体蛋白质表达变化以及群落对模型宿主细胞的反应。我们将选择牙周病原体牙龈卟啉单胞菌作为模型生物，以及具核梭菌和戈登链球菌作为牙齿生物膜中常见的生物体代表，并能够与牙龈卟啉单胞菌协同相互作用。除了这些生物体组装成群落时蛋白质表达的差异外，我们还将确定与牙龈上皮细胞接触引起的蛋白质表达的变化。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事件的基本理解。具体目标 1 单独、成对和全部定义了 P. gingivalis、S. gordonii 和 F. nucleatum 的参考蛋白质组。这将包括使用数据挖掘技术的全局蛋白质修饰分析，以及具体的全局磷酸化化学方法。我们将定义可归因于群落微生物相互作用的蛋白质相对丰度变化（相对于孤立的单个生物体）。因此，我们将建立 P. gingivalis-F 的蛋白质组。 nucleatum-S。 gordonii 群落，以及与个体蛋白质组相比对群落发展的差异反应。具体目标 2 将定义牙龈上皮细胞对 P. gingivalis-F 的影响。 nucleatum-S。戈登社区。这将涉及进行与目标 1 中相同的实验，但在上皮细胞存在的情况下，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。与目标 1 一样，这包括使用计算方法在全球范围内挖掘所有已知修饰的 PTM 数据集，此外还使用基于化学的实验方法来测量差异性整体磷酸化。在目标 3 中，根据发现的需要，我们将优先考虑目标 1 和 2 中确定的受调节蛋白，以进行额外的证实并在相应的基因中构建突变体。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。 公共卫生相关性：牙周病或牙龈疾病是发达国家牙齿脱落的主要原因，是微生物群协同活动的结果。这些生物体在组装成群落并与宿主细胞相互作用时调整其蛋白质的表达。我们将定义这些蛋白质表达变化并评估它们对疾病过程的重要性。更好地了解蛋白质变化是改进治疗和诊断策略的基本要求。