Pertussis Toxin Trafficking and Processing in Cells

百日咳毒素在细胞中的运输和加工

• 批准号: 6511602
• 负责人: NICHOLAS H CARBONETTI
• 金额: $ 29.7万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6511602
• 关键词: Bordetella pertussis; CHO cells; Golgi apparatus; MDCK cell; Saccharomyces cerevisiae; bacteria infection mechanism; bacterial vaccines; binding sites; cell membrane; cytotoxicity; endoplasmic reticulum; enzyme activity; epithelium; fluorescence microscopy; glycosylation; intracellular transport; lysine; metalloendopeptidases; pathologic process; pertussis toxin; protease inhibitor; proteolysis; serine proteinases; sulfation; transcytosis

DESCRIPTION (provided by the applicant):The short-term goal of this project is to understand the trafficking and activation of pertussis toxin (PT) within mammalian cells. Bordetella pertussis colonizes the human respiratory tract and causes the disease pertussis (whooping cough). Several systemic symptoms accompany this disease, even though B. pertussis is not thought to invade beyond the respiratory tract. Instead these symptoms are thought to be due to the action of PT, an exotoxin produced by B. pertussis. PT is an ADP ribosyltransferase that modifies several heterotrimeric G proteins, causing a wide range of effects on signaling in mammalian cells. How PT is transported within mammalian cells to arrive at its target proteins in an active form is largely unknown. In addition, how PT is transported from the respiratory tract to systemic sites is completely unknown. Understanding the mechanisms utilized by this complex toxin to achieve these effects will provide key information on the cell biology of PT and will help to provide a groundwork for studies to elucidate the role of this toxin in B. pertussis infection and disease. In addition, this information may allow development of therapeutics to combat the effects of the toxin and may also allow improvement of existing pertussis vaccines that include PT or development of novel vaccine molecules using PT as a intracellular delivery vector. We have preliminary data indicating that (i) PT may undergo retrograde intracellular trafficking through the Golgi apparatus and endoplasmic reticulum (ER) en route to its cytosolic target proteins, (ii) that proteolytic processing of the active Si subunit of cell-associated PT occurs and may be important for its activity, and (iii) that there is apparent transcytosis of active PT across intact polarized epithelial cells in culture. Therefore the specific aims of this proposal are to investigate the trafficking, processing and transcytosis of PT in mammalian cells and the key features of this toxin that mediate these events.

描述（由申请人提供）：该项目的短期目标是 了解百日咳毒素 (PT) 的运输和激活 哺乳动物细胞。百日咳博德特氏菌定植于人类呼吸道并 引起百日咳（百日咳）疾病。多种全身症状 伴随这种疾病，尽管百日咳博德特氏菌不被认为会入侵 超出呼吸道。相反，这些症状被认为是由于 PT（百日咳博德特氏菌产生的外毒素）的作用。 PT 是 ADP 核糖基转移酶可修饰多种异源三聚体 G 蛋白，从而导致 对哺乳动物细胞信号传导产生广泛的影响。 PT如何运输 在哺乳动物细胞内以活性形式到达其目标蛋白是 很大程度上不为人知。另外，PT如何从呼吸道转运 对系统部位的影响是完全未知的。了解所使用的机制 通过这种复杂的毒素来实现这些效果将提供关键信息 PT 的细胞生物学，将有助于为研究提供基础 阐明这种毒素在百日咳博德特氏菌感染和疾病中的作用。在 此外，这些信息可能有助于开发治疗方法来对抗 毒素的影响，也可能改善现有的百日咳 包括 PT 的疫苗或使用 PT 作为新型疫苗分子的开发 细胞内递送载体。我们有初步数据表明 (i) PT 可能通过高尔基体进行逆行细胞内运输 和内质网 (ER) 通往胞浆靶蛋白的途中，(ii) 细胞相关 PT 活性 Si 亚基的蛋白水解加工 发生并且可能对其活动很重要，并且（iii）有明显的 活性 PT 穿过培养物中完整极化上皮细胞的转胞吞作用。 因此，本提案的具体目标是调查 PT在哺乳动物细胞中的转运、加工和转胞吞作用及其关键 这种毒素介导这些事件的特征。