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: 8094281
• 负责人: Murray Hackett
• 金额: $ 40.47万
• 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/8094281
• 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. 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中的全球蛋白质表达变化以及社区对模型宿主细胞的反应。我们将选择作为模型生物体，将牙周病原体牙龈疟原虫以及F. nucleatum和S. gordonii作为代表牙齿生物膜中常见的生物体的代表，并能够与牙龈疟原虫协同相互作用。除了差异蛋白表达外，这些生物会将这些生物汇集到群落中，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。该研究的长期目标涉及改善我们对异性型口腔生物膜形成事件的分子水平的基本理解，以及这些生物膜与宿主细胞的相互作用。特定的目标1定义了牙龈疟原虫，戈多尼链球菌和F. nucleatum的参考蛋白质组，成对以及全部共同。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。我们将定义蛋白质的相对丰度变化（相对于孤立的单个生物），这可以归因于社区微生物相互作用。因此，我们将建立牙龈疟原虫-f的蛋白质组。核-S。与单个蛋白质组相比，Gordonii社区以及对社区发展的不同反应。特定的目标2将定义牙龈上皮细胞对牙龈斑网的影响。核-S。 Gordonii社区。这将涉及与AIM 1相同的实验，但是在上皮细胞的情况下，也具有对微生物侧的比较后翻译后修饰（PTM）分析。这将定义社区对宿主细胞存在的反应。如AIM 1所示，这包括使用计算方法为所有已知修改的PTM挖掘数据集，并使用基于化学的实验方法来测量差异全局磷酸化。在AIM 3中，我们将优先考虑AIM 1和2中确定的受调节蛋白，作为发现令，以进行附加佐证和相应基因中的突变体的构建。突变体在社区发展中的表型和对上皮细胞的反应将进一步阐明功能意义和生物学相关性。 公共卫生相关性：牙周或口香糖，是发达国家牙齿脱落的主要原因，是由微生物群体的一致活动造成的。这些生物会在聚集到群落中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贡献的重要性。对蛋白质变化的了解得到了改进，这是改善治疗和诊断策略的基本要求。