PROTEIN VIRULENCE FACTORS OF BACTERIAL PERIODONTAL PATHOGENS

细菌性牙周病原体的蛋白质毒力因子

• 批准号: 3854268
• 负责人: S LEPPLA
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3854268
• 关键词: ADP ribosylation; Bacteroides gingivalis; HIV infections; adenylate cyclase; anaerobic bacteria; bacterial proteins; bacterial toxins; cell free system; cell line; cytotoxicity; enzyme mechanism; genetic strain; human immunodeficiency virus; microorganism growth; periodontitis; reagent /indicator; virulence

A systematic and comprehensive search was begun to identify new protein virulence factors and toxins secreted by bacteria considered to be associated with periodontal disease. A collection of strains from patients with HIV and periodontal disease was sampled so as to obtain fresh disease isolates. To date, fourteen of these anaerobic species have been successfully grown and culture supernatants collected. Culture supernatants were assayed for adenylate cyclase activity; no positives were identified. Use of an internal standard of Bacillus anthracis adenylate cyclase showed that several standard media used to grow the bacterial inhibited enzyme activity. This inhibition was shown to be due to the presence of Tween 80 in the media. Media were altered to omit the detergent, thereby removing the inhibitory activity in this and potentially in other assays. Reagents and protocols were prepared for assay of the supernatants for protein tyrosine phosphatase and for ADP-ribosylation. Additional cultured cell lines were obtained that can be used for cytotoxicity tests.

开始进行系统的全面搜索以识别新蛋白质 被认为是的细菌分泌的毒力因子和毒素 与牙周疾病有关。 来自 采样了艾滋病毒和牙周疾病的患者 新鲜疾病分离株。 迄今为止，这些厌氧物种中有14种具有 成功种植并收集了培养上清液。 测定培养上清液的腺苷酸环化酶活性。不 确定了肯定。 使用杆菌的内标 炭疽蛋白腺苷酸环化酶表明，几种标准介质曾经曾经 生长细菌抑制酶活性。 显示了这种抑制作用 是由于媒体中有80分的培训。 媒体被更改为 省略洗涤剂，从而消除此中的抑制活性 可能在其他测定中。 准备试剂和协议以测定上清液的测定 蛋白酪氨酸磷酸酶和用于ADP-核糖基化。 额外的 获得可用于细胞毒性测试的培养细胞系。