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H2S production and virulence of Treponema denticola

齿垢密螺旋体的 H2S 产生和毒力

基本信息

批准号：
7161364
负责人：
LIANRUI CHU
金额：
$ 19.73万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-09-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7161364
关键词：
Abscess Address Amino Acid Sequence Ammonia Animal Model Antisense RNA Apoptosis Applications Grants Area Biochemical Biological Assay Cells Characteristics Cysteine Databases Development Environment Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Family Foundations Gamma-glutamyl transferase Gene Proteins Gene Silencing Generations Genes Genetic Glutamic Acid Glutathione Glutathione Metabolism Pathway Glycine Goals Grant Growth In Situ In Vitro Infection Investigation Lead Lesion Measures Metabolic Metabolic Pathway Metabolism Modeling Molecular Molecular Genetics Mono-S Mus Mutate N-terminal Nature Outcome Pathogenesis Pathology Pathway interactions Periodontal Diseases Periodontal Pocket Periodontium Play Production Property Proteins Pyruvate Pyruvates Recombinants Research Role Sampling Seminal Site Solid Source Substrate Interaction Sulfhydryl Compounds Sulfur Compounds Testing Tissues Translations Treponema denticola Virulence Virulence Factors Work base cystalysin cysteinylglycine design enzyme pathway enzyme substrate glutamyltransferase in vivo inhibitor/antagonist member microorganism mutant novel oral bacteria polyclonal antibody programs

项目摘要

DESCRIPTION: This application is an extension of current research examining the association between H2S production and virulence of Treponema denticola. T. denticola has been identified as an important member of a consortium of microorganisms as etiologic in the initiation and progression of periodontal diseases. In addition, the existence of volatile sulfur compounds produced at destructive sites is a characteristic feature of periodontal diseases, with H2S as a major compound in this family. Nevertheless, the metabolic pathways used to produce H2S are not well understood. Previous studies suggest that glutathione present in host cells can be a substrate for H2S production. Recently, we have shown that T. denticola has the capacity to utilize glutathione as a substrate to produce high levels of H2S. We have identified three enzymes that are required for the successful metabolism of glutathione: gamma-glutamyltransferase (GGT), cysteinylglycinase (CGase), and cystalysin. GGT converts glutathione into Cys-Gly and glutamic acid; CGase catalyze Cys-Gly to Cys and glycine; and cystalysin digests L-cysteine into H2S, ammonia, and pyruvate. We have also demonstrated that the addition of cystalysin and L-cysteine resulted in apoptosis of HGF and PDL cells and that glutathione was essential for lesion formation by T. denticola in an animal model. Based on these and other studies, three Specific Aims are proposed using biochemical, molecular genetic, and cell biologic studies to address the hypothesis that these three metabolic enzymes play a key role in T denticola pathogenesis. Specific Aim 1: To molecularly characterize the genes and proteins involved in converting glutathione to H2S. Specific Aim 2: To genetically characterize, by gene inactivation, the enzyme pathway of T. denticola that produces H2S from glutathione. Specific Aim 3: To measure the effects of the H2S/NH3 producing pathway on T. denticola virulence in vitro and in vivo. This application is designed to provide both seminal and critical information about the enzyme pathway of T. denticola to produce H2S from glutathione. The outcomes will elucidate the mechanisms of action that each member of the enzyme pathway plays in the virulence capacity of T. denticola.

产品说明：本申请是目前研究H2S生产和齿垢密螺旋体毒力之间关系的延伸。T.齿垢已被鉴定为微生物群落的重要成员，作为牙周疾病的发生和发展的病原。此外，在破坏部位产生的挥发性硫化合物的存在是牙周病的特征，H2S是该家族中的主要化合物。然而，用于产生H2S的代谢途径还不清楚。先前的研究表明，宿主细胞中存在的谷胱甘肽可以是H2S产生的底物。最近，我们证明了T.齿垢菌具有利用谷胱甘肽作为底物产生高水平H2S的能力。我们已经确定了谷胱甘肽成功代谢所需的三种酶：γ-谷氨酰转移酶（GGT）、半胱氨酰甘氨酸酶（CGase）和溶囊素。GGT将谷胱甘肽转化为Cys-Gly和谷氨酸; CGase催化Cys-Gly转化为Cys和甘氨酸;而cystalysin将L-半胱氨酸转化为H2S、氨和丙酮酸。我们还证明了加入胱溶素和L-半胱氨酸可导致HGF和PDL细胞凋亡，谷胱甘肽是T.在动物模型中的牙垢。基于这些和其他研究，提出了三个具体目标，使用生物化学，分子遗传学和细胞生物学研究，以解决这三种代谢酶在齿垢锥虫发病机制中发挥关键作用的假设。具体目标1：对谷胱甘肽转化为H2S过程中涉及的基因和蛋白质进行分子表征。具体目标2：通过基因失活，对T.由谷胱甘肽产生硫化氢的齿垢菌。具体目标3：测量H2S/NH3产生途径对T.在体外和体内的齿垢毒力。这个应用程序的目的是提供有关T。从谷胱甘肽中产生H2S。这些结果将阐明酶途径的每个成员在T.齿垢