A chemical proteomics survey of Plasmodium gametocyte development

疟原虫配子细胞发育的化学蛋白质组学调查

• 批准号: 9537357
• 负责人: Christoph Grundner
• 金额: $ 4.33万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-23 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9537357
• 关键词: Anopheles Genus; Antimalarials; Base Sequence; Biological Assay; Biology; Blood; Chemicals; Culicidae; Data; Development; Disease; Drug Targeting; Enzymes; Family; Female; Frequencies; Human; Individual; Infection; Label; Lipase; Malaria; Mass Spectrum Analysis; Measures; N-terminal; Parasites; Partner in relationship; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Physiological; Plasmodium; Plasmodium falciparum; Play; Prevention; Proteins; Proteome; Proteomics; Recombinants; Role; Serine Hydrolase; Serine Protease; Slice; Surveys; activity-based protein profiling; asexual; base; enzyme activity; esterase; male; member; new therapeutic target; novel strategies; prevent; tool; transmission process; vector mosquito; virtual

Abstract The spread of malaria parasites by the Anopheles mosquito vector sustains high levels of reinfection in endemic regions and severely impedes malaria control. As a result, efforts to eliminate malaria must include strategies that prevent parasite transmission. Because only the asexual blood stages of the parasite cause disease, they are the main target of most antimalarials. In contrast, the developmental stage responsible for transmission, the gametocyte, is only poorly understood, and most drugs that are effective against the asexual blood stages are ineffective against gametocytes. Here, we will directly measure and identify the activity of a particularly druggable enzyme family, the serine hydrolases, in the native proteome of Plasmodium falciparum (Pf) schizont and gametocyte stages. We will use activity-based protein profiling (ABPP) with a serine hydrolase-directed activity probe in combination with quantitative mass spectrometry to survey a broad cross- section of enzyme activity during the transition from the asexual to the sexual, mosquito-transmissible form. Because ABPP is a mechanism-based and sequence-independent approach, these studies will also capture the activity of Pf proteins with unknown function. We will further identify the serine proteases among the hits, and define the physiologic substrates of known and previously unknown proteases. Defining the SH activity changes associated with the transition from asexual to sexual Pf gametocytes will provide a new and broad activity-based view of this developmental transition, identify potential transmission blocking targets, and provide experimental annotation for a large number of proteins with unknown function.

抽象的 按蚊媒介传播疟原虫导致疟疾的再感染率很高 流行地区并严重阻碍疟疾控制。因此，消除疟疾的努力必须包括 防止寄生虫传播的策略。因为只有寄生虫的无性血液阶段才会导致 疾病，它们是大多数抗疟药的主要目标。相反，发育阶段负责 人们对配子体这一传播途径知之甚少，大多数对无性生殖有效的药物 血液阶段对配子体无效。在这里，我们将直接测量和识别 恶性疟原虫天然蛋白质组中特别可药物化的酶家族丝氨酸水解酶 (Pf) 裂殖体和配子体阶段。我们将使用基于活性的蛋白质分析 (ABPP) 和丝氨酸 水解酶定向活性探针与定量质谱相结合，可调查广泛的交叉 从无性到有性、蚊子传播形式转变过程中酶活性的一部分。 由于 ABPP 是一种基于机制且与序列无关的方法，因此这些研究还将捕获 功能未知的 Pf 蛋白的活性。我们将进一步鉴定命中中的丝氨酸蛋白酶， 并定义已知和以前未知的蛋白酶的生理底物。定义 SH 活动 与从无性到有性 Pf 配子细胞的转变相关的变化将提供新的、广泛的 基于活动的观点来看待这一发展转变，确定潜在的传播阻断目标，以及 为大量功能未知的蛋白质提供实验注释。