Molecular mechanisms of Plasmodium fertilization

疟原虫受精的分子机制

• 批准号: 10064068
• 负责人: MARCELO JACOBS-LORENA
• 金额: $ 40.94万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-07 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064068
• 关键词: Affinity; Antibodies; Antibody Formation; Binding; Biological Assay; Biology; Biotin; Blood; Cell Separation; Communicable Diseases; Culicidae; Development; Disease; Economics; Enzyme-Linked Immunosorbent Assay; Event; Female; Fertilization; Genes; Germ Cells; Goals; Human; Immunofluorescence Immunologic; In Vitro; Intervention; Knock-out; Knowledge; Laboratories; Lead; Libraries; Life Cycle Stages; Ligands; Light; Liver; Malaria; Mass Spectrum Analysis; Membrane Proteins; Midgut; Molecular; Molecular and Cellular Biology; Nature; Organism; Parasites; Parasitic Diseases; Peptide Phage Display Library; Peptide antibodies; Peptides; Persons; Phage Display; Plasmodium; Play; Population; Process; Proteins; Proteome; Receptor Gene; Recombinants; Refractory; Research; Role; Salivary Glands; Sexual Development; Sexual Reproduction; Structure; Surface; Surface Plasmon Resonance; Tertiary Protein Structure; Testing; Transgenic Organisms; Western Blotting; base; experimental study; in vivo; insight; malaria transmission; male; novel; parasite invasion; peptide structure; peptidomimetics; prevent; receptor; receptor function; sex; tool; transmission process; transmission-blocking vaccine; vector mosquito; Affinity; Antibodies; Antibody Formation; Binding; Biological Assay; Biology; Biotin; Blood; Cell Separation; Communicable Diseases; Culicidae; Development; Disease; Economics; Enzyme-Linked Immunosorbent Assay; Event; Female; Fertilization; Genes; Germ Cells; Goals; Human; Immunofluorescence Immunologic; In Vitro; Intervention; Knock-out; Knowledge; Laboratories; Lead; Libraries; Life Cycle Stages; Ligands; Light; Liver; Malaria; Mass Spectrum Analysis; Membrane Proteins; Midgut; Molecular; Molecular and Cellular Biology; Nature; Organism; Parasites; Parasitic Diseases; Peptide Phage Display Library; Peptide antibodies; Peptides; Persons; Phage Display; Plasmodium; Play; Population; Process; Proteins; Proteome; Receptor Gene; Recombinants; Refractory; Research; Role; Salivary Glands; Sexual Development; Sexual Reproduction; Structure; Surface; Surface Plasmon Resonance; Tertiary Protein Structure; Testing; Transgenic Organisms; Western Blotting; base; experimental study; in vivo; insight; malaria transmission; male; novel; parasite invasion; peptide structure; peptidomimetics; prevent; receptor; receptor function; sex; tool; transmission process; transmission-blocking vaccine; vector mosquito

ABSTRACT Malaria remains one of the most devastating infectious diseases. It kills over a million people every year while causing immense suffering and economic losses worldwide. Whereas much progress has been made in understanding the life cycle of the parasite in the human host and in the mosquito vector, significant gaps of knowledge remain. Fertilization of malaria parasites is a poorly understood process that takes place in the lumen of the mosquito gut. This process is important because survival in nature is completely dependent on the ability of the parasite to undergo sexual reproduction. The proposed research aims to identify molecular interactions that take place during fertilization of malaria parasites. This project is based on an unorthodox approach (identification of peptides that bind to the gamete surface) made possible by the recent development of an important tool, a transgenic parasite that produces red-fluorescent female gametes and green-fluorescent male gametes. Pure populations of malaria female and male gametes from this transgenic parasite were isolated by cell sorting and then used to screen a phage display library for peptides that recognize molecules on the gamete surfaces. A peptide (FG1) that binds to female gametes and another peptide (MG1) that binds to male gametes were identified. Importantly, when added to a malaria infectious blood meal, each of these peptides blocked parasite fertilization, suggesting that the peptides bound to a receptor and prevented its interaction with a ligand on the gamete of the opposite sex. Our working hypothesis is that the peptide structurally mimics the gamete ligand and that peptide and ligand compete for binding to the corresponding receptor. This proposal lays out a research plan to identify the receptor to which the FG1 and MG1 peptides bind and the ligands on the gametes of the opposite sex that the two peptides structurally mimic. An additional aim is to characterize the proteome on the surface of female gametes before and after fertilization, to gain insights on additional proteins involved in fertilization and possible block to polyspermy. Elucidation of mechanisms of fertilization is important not only for understanding the basic biology of malaria and other parasitic diseases but could also lead to the identification of new targets for blocking transmission and the spread of disease. Moreover, should the mechanisms be conserved, our findings could be extended to the biology of fertilization of higher organisms.

抽象的 疟疾仍然是最具破坏性的传染病之一。它杀死了超过一百万的人 每年都会在全球造成巨大的痛苦和经济损失。而很多 在理解人类宿主和在 蚊子向量，知识的差距仍然存在。疟疾寄生虫的受精是 在蚊子肠道内发生的鲜为人知的过程。这个过程是 重要，因为自然界的生存完全取决于寄生虫的能力 进行有性繁殖。 拟议的研究旨在确定受精过程中发生的分子相互作用 疟疾寄生虫。该项目基于一种非正统的方法（肽的识别 与配子表面结合的）是由于最近开发重要工具而成为可能的 一种产生红荧光女配子和绿色男性的转基因寄生虫 配子。来自这种转基因寄生虫的疟疾雌性和男配子的纯种群 通过细胞分类隔离，然后用于筛选噬菌体显示库的肽 识别配子表面上的分子。与女配子结合的肽（FG1） 鉴定出与男配子结合的另一个肽（MG1）。重要的是，添加时 疟疾感染性血液粉，这些肽中的每一个都阻塞了寄生虫受精， 提示肽结合到受体并防止其与配体上的相互作用 异性的配子。我们的工作假设是肽在结构上模仿 配子配体以及该肽和配体竞争与相应的结合 受体。该建议制定了一项研究计划，以识别FG1和 MG1肽结合和在异性配子上的配体这两种肽 结构模仿。另一个目的是表征女性表面上的蛋白质组 施肥前后的配子，以了解有关涉及的其他蛋白质的见解 受精，并可能阻止多工。 阐明受精机制不仅对理解基本 疟疾和其他寄生疾病的生物学，但也可能导致对新的鉴定 阻止传播和疾病传播的目标。而且，应有机制 保守，我们的发现可以扩展到较高生物体受精的生物学。