Structure and dynamics of bacterial membrane protein - receptor interactions

描述（由申请人提供）：许多专性细菌膜蛋白通过模仿或操纵宿主机制来劫持人类细胞通路。本研究的目的是研究细菌外膜蛋白的结构和动力学及其与宿主受体的相互作用。具体而言，研究集中在来自淋病奈瑟菌和脑膜炎奈瑟菌的外膜不透明相关蛋白（Opa）上，其通过与宿主受体结合而诱导细菌在非吞噬细胞中的吞噬。Opa蛋白结合各种宿主受体，并基于宿主受体选择性分为两个家族。较大类别OpaCEA与癌胚抗原样细胞粘附分子（CEACAMs）结合，较小类别OpaHS与两种不同受体结合;硫酸乙酰肝素蛋白聚糖受体（HSPG）通过肝素介导的与纤连蛋白或纤连蛋白的相互作用直接和间接与整联蛋白受体结合。Opa蛋白是完整的外膜蛋白，并预测具有八链2桶折叠。两个细胞外环（HV 1和HV 2）在Opa蛋白之间具有最多的序列变异，并决定宿主受体特异性。不仅HV环区分HSPG和CEACAM受体，而且OpaCEA蛋白可以基于与七种CEACAM受体中的四种的选择性结合而进一步分为亚组。使用核磁共振、电子顺磁共振、等温滴定量热法和突变，将确定这些相互作用的分子决定因素。这些结果将为淋病奈瑟菌和脑膜炎奈瑟菌的发病机制提供深入的了解，因此，合理设计新型抗生素的潜力。此外，重构的Opa蛋白可用于疫苗开发。然而，这项研究的最新应用在于Opa蛋白通过三种不同机制特异性靶向宿主受体以诱导非吞噬细胞中的内吞作用的能力。这种能力可能是有用的脂质体药物载体。脂质体包封的治疗剂（例如酶、抑制剂和肽）选择性地进入活细胞和可能的组织的细胞质的潜在能力对于许多疾病的治疗至关重要。了解这三种Opa介导的进入机制的分子决定因素可能有助于脂质体递送机制的发展。

DESCRIPTION (provided by applicant): Many obligate bacterial membrane proteins hijack human cellular pathways by mimicking or manipulating host machinery. The goal of this research is to investigate the structure and dynamics of bacterial outer membrane proteins and their interactions with host receptors. Specifically, research is focused on the outer membrane opacity-associated proteins (Opa) from Neisseria gonorrhoeae and Neisseria meningitides, which induce engulfment of the bacterium in non-phagocytic cells by binding to host receptors. Opa proteins bind to various host receptors and are classified into two families based on host receptor selectivity. The larger class, OpaCEA, bind to carcinoembryonic antigen-like cellular adhesion molecules (CEACAMs), and the smaller class, OpaHS, bind to two different receptors; the heparansulfate proteoglycan receptors (HSPGs) directly and indirectly to integrin receptors via a heparin- mediated interaction with fibronectin or vibironectin. Opa proteins are integral outer membrane proteins and predicted to have an eight-stranded 2-barrel fold. Two of the extracellular loops (HV1 and HV2) have the most sequence variation between Opa proteins and determine the host receptors specificity. Not only do the HV loops discriminate between HSPG and CEACAM receptors, but OpaCEA proteins can be further divided into subgroups based on the selective binding to four of the seven CEACAM receptors. Using nuclear magnetic resonance, electron paramagnetic resonance, isothermal titration calorimetry, and mutagenesis, the molecular determinants of these interactions will be determined. The results will provide insight into the pathogenesis of Neisseria gonorrhoeae and Neisseria meningitides and, therefore, the potential for the rational design of novel antibiotics. In addition, the reconstituted Opa proteins may be useful for vaccine development. However, the most novel application of this research lies in the ability of Opa proteins to target host receptors specifically via three different mechanisms to induce endocytosis in non-phagocytic cells. This ability may be useful for liposome pharmaceutical carriers. The potential ability of liposome encapsulated therapeutics (e.g. enzymes, inhibitors, and peptides) to enter the cytoplasm of living cells and possibly tissue selectively is of crucial importance to the treatment of many diseases. Understanding the molecular determinants of the three Opa-mediated entry mechanisms may facilitate the development of liposome delivery mechanisms.