ELUCIDATING & DISRUPTING PLASMODIUM ERYTHROCYTE-BINDING LIKE LIGAND INTERACTIONS

摘要 对于疟原虫侵入红细胞所需的相互作用的理解存在根本性的空白 寄生虫我们的长期目标是确定红细胞侵袭期间受体-配体相互作用的方式 在分子水平上介导，以及如何利用它们进行预防，治疗和诊断 目的本申请的目的是了解受体识别的分子基础， 红细胞结合样（EBL）家族，并建立中和性单克隆抗体的抑制模式， 针对这个家族的抗体本申请的中心假设是，结合结构域（DBL EBL家族成员的结构域）具有二聚化的保守折叠，产生受体结合口袋 以及用于识别各种受体的通道，以及抑制性抗体靶向结合的通道。 口袋和渠道，无论是通过防止其形成或其可达性的理由，建议 研究表明，一旦确定了EBL家族成员的功能区域， 用于疫苗设计、基于蛋白质的新型治疗和/或诊断。因此，拟议的 研究与国家卫生研究院的使命有关，即开发基础知识， 人类疾病的负担。此外，拟议的研究将促进我们对受体的理解- 配体相互作用、配体-抗体相互作用和微生物发病机制。支持强大的初步 数据，这一假设将通过追求两个特定的目标进行测试：1）确定受体结合口袋的 EBL家族成员;和2）确定中和抗体如何结合DBL结构域并抑制 入侵在第一个目标下，将使用结构、体外和体内研究来全面确定 EBL家族介导的相互作用的分子细节。在第二个目标下， 相互作用作图研究将揭示EBL家族的表位，这些表位被中和抗体靶向 鉴定具有最大中和潜力的表位。这项研究意义重大，因为 它有望推进和扩展我们对受体-配体和配体-抗体相互作用的理解 并提供发展诊断、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.