ELUCIDATING & DISRUPTING PLASMODIUM ERYTHROCYTE-BINDING LIKE LIGAND INTERACTIONS

摘要 对疟原虫侵袭红细胞所需的相互作用的理解存在着根本的差距 寄生虫。我们的长期目标是确定在红细胞入侵过程中受体-配体的相互作用如何 在分子水平上的调节以及如何利用它们进行预防、治疗和诊断 目的。本应用程序的目的是通过以下方式了解受体识别的分子基础 红细胞结合样蛋白(EBL)家族及中和单抗抑制模式的建立 针对这个家族的抗体。该应用程序的中心假设是结合结构域(DBL EBL家族成员的结构域)具有一个保守的折叠，该折叠可以二聚化，产生受体结合口袋 以及用于识别各种受体的通道，以及以结合为目标的抑制性抗体 口袋和渠道，防止它们的形成或进入，拟议的理由是 研究表明，一旦确定了EBL家族成员的功能区，这些区域就可以 用于疫苗设计、新的基于蛋白质的疗法和/或诊断。因此，拟议的 研究与美国国立卫生研究院发展基础知识的使命部分相关，这些基础知识可能会减少 人类疾病的负担。此外，拟议的研究将促进我们对受体- 配体相互作用、配体-抗体相互作用和微生物发病机制。有强劲的初步支持 数据，这一假说将通过追求两个特定的目标来检验：1)识别受体结合口袋 EBL家族成员；以及2)确定中和抗体如何与DBL结构域结合并抑制 入侵。在第一个目标下，将使用结构、体外和体内研究来综合确定 EBL家族介导的相互作用的分子细节。在第二个目标下，结构性和 相互作图研究将揭示EBL家族的表位，以中和抗体为靶点 确定具有最大中和潜力的表位。这项拟议的研究具有重要意义，因为 这有望促进和扩大我们对受体-配体和配体-抗体相互作用的理解。 并提供制定诊断、预防和治疗干预措施所需的知识 疟疾。

ABSTRACT There is a fundamental gap in understanding of interactions required for erythrocyte invasion by Plasmodium parasites. Our long-term goal is to determine how receptor-ligand interactions during erythrocyte invasion are mediated at the molecular level and how they can be exploited for preventative, therapeutic and diagnostic purposes. The objectives of this application are to understand the molecular basis of receptor recognition by the Erythrocyte-binding like (EBL) family and to establish the mode of inhibition of neutralizing monoclonal antibodies that target this family. The central hypothesis of the application is that the binding domains (DBL domains) of EBL family members possess a conserved fold that dimerizes, creating receptor-binding pockets and channels that are used to recognize a variety of receptors, and that inhibitory antibodies target binding pockets and channels either by preventing their formation or their accessibility The rationale for the proposed research is that once the functional regions of the EBL family members have been determined these can be exploited for vaccine design, novel protein-based therapeutics and/or diagnostics. Thus, the proposed research is relevant to that part of the NIH's mission of developing fundamental knowledge that may reduce the burden of human disease. In addition, the proposed research will advance our understanding of receptor- ligand interactions, ligand-antibody interactions and microbial pathogenesis. Supported by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Identify the receptor binding pockets of the EBL family members; and 2) Determine how neutralizing antibodies bind to DBL domains and inhibit invasion. Under the first aim, structural, in vitro and in vivo studies will be used to comprehensively determine the molecular details of interactions mediated by the EBL family. Under the second aim, structural and interaction mapping studies will reveal epitopes of the EBL family targeted by neutralizing antibodies towards identifying epitopes that have the greatest neutralizing potential. The proposed research is significant because it is expected to advance and expand our understanding of receptor-ligand and ligand-antibody interactions and to provide the knowledge required to develop diagnostics, preventative and therapeutic interventions for malaria.