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.

项目摘要 /摘要 疟疾折磨了数亿，每人杀死60万人 一年，主要是撒哈拉以南非洲的儿童。天冬氨酸长期以来一直是 感兴趣的抗疟疾目标。大量天冬氨酸蛋白酶抑制剂 已经开发了反对文化中寄生虫的有力，但它们的具体 目标是难以捉摸的。纤溶酶X（PMX）是最不明显的之一 在无性炎性疟疾寄生虫中发现的天冬氨酸蛋白酶。我们有 最近发现，PMX是促进性促寄生虫出口的关键酶，并且 入侵。它激活主触发类蛋白酶样蛋白酶1（sub1）， 发起蛋白水解事件，使梅罗龙夫人能够从宿主的红血中脱颖而出 细胞（RBC）并入侵新鲜的RBC。我们已经确定了一类天冬氨酸 蛋白酶抑制剂称为氨基氢剂，似乎通过 PMX封锁，防止SUB1激活和损害出口/入侵。 PMX 敲低的抑制剂作用。其中一种抑制剂有利 药代动力学特性并在啮齿动物疟疾模型中提供口服治愈。我们 相信PMX是一个令人兴奋的新药目标，但需要更好地描述其 功能以告知正在进行的药物开发并增强我们对 寄生虫生物学。 为了解决这些问题，AIM 1将检查PMX的特殊性，并将 解决sub1成熟PMX是否直接的问题。生化 使用SUB1用作底物的孤立PMX和随机肽的测定 库将执行。 AIM 2将重点放在PMX相互作用上。是sub1 唯一的底物？分泌的囊泡中还有什么称为外部， PMX和SUB1都居住在哪里？ AIM 3将解决PMX的问题 本身被激活。我们的初步数据表明必须有一个 上游酶。我们将表征加工并寻找成熟酶。 我们预计拟议的研究将对 疟疾的发病机理，并将指向新疗法的道路 毁灭性疾病。

项目成果

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

• DOI: 10.1038/s41467-022-32271-7
• 发表时间: 2022-08-04
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Mukherjee, Sumit;Nguyen, Suong;Sharma, Eashan;Goldberg, Daniel E.
• 通讯作者: Goldberg, Daniel E.

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

• DOI: 10.1128/mbio.00673-23
• 发表时间: 2023-04-25
• 期刊: MBIO
• 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.

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

• DOI: 10.1111/cmi.12868
• 发表时间: 2018-10
• 期刊: Cellular microbiology
• 影响因子: 3.4
• 作者: Glushakova S;Beck JR;Garten M;Busse BL;Nasamu AS;Tenkova-Heuser T;Heuser J;Goldberg DE;Zimmerberg J
• 通讯作者: Zimmerberg J

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。

• DOI: 10.1021/acsinfecdis.8b00197
• 发表时间: 2019
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Singh,Snigdha;Rajendran,Vinoth;He,Jiang;Singh,AmitK;Achieng,AngelaO;Vandana;Pant,Akansha;Nasamu,ArmiyawS;Pandit,Mansi;Singh,Jyoti;Quadiri,Afshana;Gupta,Nikesh;Poonam;Ghosh,PrahladC;Singh,BrajendraK;Narayanan,Latha;Kempai
• 通讯作者: Kempai