Plasmepsin X function in Plasmodium

Plasmodium 中 Plasmepsin X 的功能

• 批准号: 10322714
• 负责人: Daniel E. Goldberg
• 金额: $ 38.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-02 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322714
• 关键词: Address; Africa South of the Sahara; Antimalarials; Aspartic Endopeptidases; Back; Binding; Biochemical; Biological Assay; Biology; Biotinylation; Child; Cysteine; Data; Disease; Drug Kinetics; Enzymes; Erythrocytes; Event; Immunoprecipitation; Impairment; In Vitro; Invaded; Label; Lead; Malaria; Mass Spectrum Analysis; Modeling; Mutate; Oral; Organelles; Parasites; Pathogenesis; Peptide Hydrolases; Persons; Phenocopy; Physiologic pulse; Plasmodium; Process; Property; Protease Inhibitor; Proteins; Proteolysis; Proteomics; Random Peptide Libraries; Recombinants; Rodent; Secretory Vesicles; Serine Protease; Site; Specificity; Substrate Specificity; Synthetic Peptide Libraries; Time; asexual; drug development; inhibitor; insight; interest; knock-down; mutant; new therapeutic target; novel; novel therapeutics; plasmepsin; prevent

PROJECT SUMMARY / ABSTRACT Malaria afflicts several hundred million and kills more than 600,000 people each year, mostly children in Sub-Saharan Africa. Aspartic proteases have long been antimalarial targets of interest. A large number of aspartic protease inhibitors that are potent against parasites in culture have been developed, but their specific targets have been elusive. Plasmepsin X (PMX) is one of the least characterized aspartic proteases found in asexual intraerythrocytic malaria parasites. We have recently found that PMX is a key enzyme for intraerythrocytic parasite egress and invasion. It activates the master trigger subtilysin-like protease 1 (SUB1), launching proteolytic events that allow merozoites to get out of the host red blood cell (RBC) and invade fresh RBCs. We have identified a class of aspartic protease inhibitors called aminohydantoins that appear to kill parasites through PMX blockade, preventing SUB1 activation and impairing egress/invasion. PMX knockdown phenocopies inhibitor action. One of the inhibitors has favorable pharmacokinetic properties and gives oral cure in a rodent malaria model. We believe that PMX is an exciting new drug target but need to better characterize its function to inform ongoing drug development and enhance our understanding of parasite biology. To address these questions, aim 1 will examine the specificity of PMX and will address the question of whether SUB1 maturation by PMX is direct. Biochemical assays using isolated PMX with SUB1 as a substrate and with a random peptide library will be performed. Aim 2 will focus on what PMX interacts with. Is SUB1 the only substrate? What else is in the secretory vesicle called the exoneme, where PMX and SUB1 both reside? Aim 3 will address the question of how PMX itself gets activated. Our preliminary data suggest that there must be an upstream enzyme. We will characterize the processing and look for a maturase. We anticipate that the proposed studies will yield great insight into the pathogenesis of malaria and will point the way to new therapies for this devastating disease.

项目概要/摘要 疟疾困扰着数亿人并导致超过 60 万人死亡 年，主要是撒哈拉以南非洲地区的儿童。天冬氨酸蛋白酶长期以来 感兴趣的抗疟靶点。大量的天冬氨酸蛋白酶抑制剂 已开发出对文化中的寄生虫有效的药物，但其具体 目标一直难以捉摸。 Plasmepsin X (PMX) 是特征最少的酶之一 在无性红细胞内疟原虫中发现的天冬氨酸蛋白酶。我们有 最近发现 PMX 是红细胞内寄生虫排出的关键酶 入侵。它激活主触发枯草溶菌素样蛋白酶 1 (SUB1)， 启动蛋白水解事件，使裂殖子脱离宿主红血 细胞（红细胞）并侵入新鲜红细胞。我们已经确定了一类天冬氨酸 称为氨基乙内酰脲的蛋白酶抑制剂似乎可以通过以下方式杀死寄生虫 PMX 封锁，防止 SUB1 激活并削弱出口/入侵。聚甲基丙烯酸甲酯 敲低表型抑制剂的作用。其中一种抑制剂具有良好的 药代动力学特性，并在啮齿动物疟疾模型中提供口服治疗。我们 相信 PMX 是一个令人兴奋的新药物靶点，但需要更好地描述其特征 为正在进行的药物开发提供信息并增强我们的理解 寄生虫生物学。 为了解决这些问题，目标 1 将检查 PMX 的特殊性，并将 解决 PMX 是否直接使 SUB1 成熟的问题。生化 使用分离的 PMX（以 SUB1 作为底物和随机肽）进行测定 库将被执行。目标 2 将重点关注 PMX 与之交互的内容。是SUB1 唯一的基材？分泌囊泡中还有什么叫做外显子， PMX 和 SUB1 都驻留在哪里？目标 3 将解决 PMX 如何 本身被激活。我们的初步数据表明，必须有一个 上游酶。我们将表征加工过程并寻找成熟酶。 我们预计拟议的研究将对 疟疾的发病机制，并将为该疾病的新疗法指明道路 毁灭性的疾病。

项目成果

Activation of the Plasmodium Egress Effector Subtilisin-Like Protease 1 Is Mediated by Plasmepsin X Destruction of the Prodomain.

疟原虫出口效应器枯草杆菌蛋白酶 1 的激活是由 Plasmepsin X 对前结构域的破坏介导的。

• 发表时间: 2023-04-25
• 影响因子: 6.4
• 作者: Mukherjee, Sumit;Nasamu, Armiyaw S;Rubiano, Kelly C;Goldberg, Daniel E
• 通讯作者: Goldberg, Daniel E

Malaria parasite plasmepsins: More than just plain old degradative pepsins.

疟疾寄生虫血浆蛋白酶：不仅仅是普通的老式降解胃蛋白酶。

• 发表时间: 2020
• 作者: Nasamu, Armiyaw S;Polino, Alexander J;Istvan, Eva S;Goldberg, Daniel E
• 通讯作者: Goldberg, Daniel E

Fast-Acting Small Molecules Targeting Malarial Aspartyl Proteases, Plasmepsins, Inhibit Malaria Infection at Multiple Life Stages.

针对疟疾天冬氨酰蛋白酶、纤溶酶的速效小分子可抑制多个生命阶段的疟疾感染。

• 发表时间: 2019
• 影响因子: 5.3
• 作者: Singh, Snigdha;Rajendran, Vinoth;He, Jiang;Singh, Amit K;Achieng, Angela O;Vandana;Pant, Akansha;Nasamu, Armiyaw S;Pandit, Mansi;Singh, Jyoti;Quadiri, Afshana;Gupta, Nikesh;Poonam;Ghosh, Prahlad C;Singh, Brajendra K;Narayanan, Latha;Kempai
• 通讯作者: Kempai

Maturation and substrate processing topography of the Plasmodium falciparum invasion/egress protease plasmepsin X.

恶性疟原虫侵入/流出蛋白酶plasmepsin X的成熟和底物加工形貌。

• 发表时间: 2022-08-04
• 影响因子: 16.6
• 作者: Mukherjee, Sumit;Nguyen, Suong;Sharma, Eashan;Goldberg, Daniel E
• 通讯作者: Goldberg, Daniel E

Rounding precedes rupture and breakdown of vacuolar membranes minutes before malaria parasite egress from erythrocytes.

在疟原虫从红细胞中排出之前几分钟，液泡膜就会发生破裂和破裂。

• 发表时间: 2018
• 影响因子: 3.4
• 作者: Glushakova, Svetlana;Beck, Josh R;Garten, Matthias;Busse, Brad L;Nasamu, Armiyaw S;Tenkova;Heuser, John;Goldberg, Daniel E;Zimmerberg, Joshua
• 通讯作者: Zimmerberg, Joshua