Analysis of newly identified adhesin used by pathogenic Gram-negative bacteria

致病性革兰氏阴性菌使用的新发现的粘附素的分析

• 批准号: 8518227
• 负责人: Kim Orth
• 金额: $ 22.42万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8518227
• 关键词: Actins; Adhesions; Animals; Attenuated; Bacteria; Bacterial Adhesins; Bacterial Infections; Bacterial Typing; Binding; Biochemical; Biological; Cell Adhesion Molecules; Cell-Matrix Junction; Cells; Consumption; Cytoskeleton; Fibronectin Receptors; Fibronectins; Foundations; Future; Gastroenteritis; Genes; Goals; Gram-Negative Bacteria; Hemolysin; Infection; Investigation; Lead; Lipids; Mediating; Membrane Proteins; Phosphatidic Acid; Production; Proteins; Receptor Cell; Recombinants; Role; Seafood; Signal Pathway; Signal Transduction; Staging; Surface; Type III Secretion System Pathway; Vibrio parahaemolyticus; cytotoxicity; inhibitor/antagonist; insight; interest; novel; pathogen; protein protein interaction; receptor; research study

DESCRIPTION (provided by applicant): The aim of this proposal is to characterize a newly multivalent adhesion molecule (MAM7) found in the majority of Gram-negative pathogenic animal bacteria. We propose that MAM7 is involved in the initial contact of bacterial pathogens with host cells. As a representative for this type of molecule, we will use the MAM7 protein from Vibrio parahaemolyticus, an emerging pathogen that causes gastroenteritis through consumption of raw or undercooked seafood, to study the multivalent binding to host cells. We observe MAM7 dependent changes in the actin cytoskeleton in host cells. We predict that binding of MAM7 will trigger the production of more adhesion factors in the bacterial pathogens. Thus as one of our aims, we would like to uncover the signaling machinery activated in the bacteria and the host cell upon binding of MAM7. In another aim, we would like to understand the biochemical interaction of MAM7 with its two host cell receptors, fibronectin and phosphatidic acid. This information will be extremely valuable for future studies focused on MAM7 inhibitors. Finally, we observe that binding of non-pathogenic bacteria expressing MAM7 (BL21-MAM7) to host cells ameliorates cytotoxicity of Gram-negative pathogens expressing MAM7. We are interested in determining how broadly MAM7 provides a competitive advantage over other types of bacterial pathogens that do not express MAM7 but other types of adhesins. Together, these experiments will allow us to characterize the newly identified multivalent adhesion molecule MAM7 and will help to elucidate its value as a competitive inhibitor for a wide variety of bacterial pathogens.

描述（由申请人提供）：该提案的目的是表征大多数革兰氏阴性致病动物细菌中新近多价粘附分子（MAM7）。我们建议MAM7参与细菌病原体与宿主细胞的初始接触。作为这种类型分子的代表性，我们将使用来自Vibrio Parahayticus的MAM7蛋白，这是一种新兴的病原体，通过消耗原始或未煮熟的海鲜引起胃肠炎，研究与宿主细胞的多价结合。我们观察到宿主细胞中肌动蛋白细胞骨架的MAM7依赖性变化。我们预测MAM7的结合将触发细菌病原体中更多粘附因子的产生。因此，作为我们的目标之一，我们希望在MAM7结合后发现在细菌和宿主细胞中激活的信号机制。在另一个目的中，我们想了解MAM7与其两个宿主细胞受体的生化相互作用：纤连蛋白和磷脂酸。对于关注MAM7抑制剂的未来研究，此信息将非常有价值。最后，我们观察到表达MAM7（BL21-MAM7）宿主细胞的非致病细菌的结合可以改善表达MAM7的革兰氏阴性病原体的细胞毒性。我们有兴趣确定与其他类型的细菌病原体相比，MAM7如何提供竞争优势，这些细菌病原体不会表达MAM7而不是其他类型的粘附素。总之，这些实验将使我们能够表征新鉴定的多价粘附分子MAM7，并有助于阐明其作为多种细菌病原体的竞争抑制剂的价值。