Structure and function of the Plasmodium myosin XIV-actin glideosome.

疟原虫肌球蛋白 XIV-肌动蛋白滑胶体的结构和功能。

• 批准号: 10650841
• 负责人: KATHLEEN M TRYBUS
• 金额: $ 66.16万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-11 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10650841
• 关键词: ATP phosphohydrolase; Actins; Actomyosin; Africa; Age; Artemisinins; Binding; Binding Sites; Biological; Biological Assay; Cell division; Cell physiology; Cells; Cellular biology; Cessation of life; Child; Complex; Cryoelectron Microscopy; Crystallization; Culicidae; Development; Drug resistance; Erythrocytes; Foundations; Future; Genetic; Genetic study; Goals; Heart; Human; In Vitro; Invaded; Investigation; Kinetics; Licensure; Life Cycle Stages; Light; Malaria; Malaria Vaccines; Molecular; Motion; Motor; Myosin ATPase; Myosin Type V; N-terminal; Organelles; Parasites; Parasitic infection; Pathogenesis; Pharmaceutical Preparations; Phosphorylation; Phosphorylation Site; Plasmodium; Plasmodium falciparum; Play; Positioning Attribute; Power stroke; Process; Property; Reporting; Research; Role; Sexual Development; Sporozoites; Structure; Tissues; Vaccines; Virulent; Work; X-Ray Crystallography; cell motility; conditional knockout; druggable target; global health; imaging study; in vivo; inhibitor; insight; live cell imaging; malaria infection; mutant; myosin VI; new therapeutic target; novel; optic trap; optical traps; small molecule inhibitor; superresolution imaging; tool; trafficking; virtual

Malaria infection in humans, caused by single-celled parasites from the genus Plasmodium, is a major global health challenge. Despite marked progress in the last 15 years, more than 400 million deaths occur worldwide annually, the majority being children under age 5. Recent licensure of the first ever malaria vaccine heralds a new era in efforts to control malaria, but the relatively modest efficacy of the RTS,S vaccine means that complementary approaches will be essential if the WHO's goal of a 90% reduction in rates by 2030 is to be realized. Malaria parasites are motile throughout their complex human and mosquito lifecycle. They move by a process called gliding motility, which underpins their ability to reach, cross, and enter host tissues and cells. Gliding is powered by a parasite actomyosin motor the disruption of which kills the infectious parasite. Towards development of the parasite actomyosin motor as a druggable target, our collaborative team has worked to characterize the essential class XIV single-headed myosin motor PfMyoA, the core of gliding motility. We were the first to characterize and crystallize PfMyoA, demonstrating that its function is uniquely tuned by N-terminal heavy chain phosphorylation. We were the first to show the essential role of PfMyoA and its essential light chain in powering red blood cell (RBC) invasion, the stage responsible for all malaria pathogenesis. We have since used PfMyoA mutants to reveal the energetic barriers necessary for RBC invasion using live cell imaging. These foundations expertly position our team to extend investigation of gliding motility across the malaria lifecycle and explore additional Plasmodium myosins and their cellular roles, which are the combined aims of this competitive renewal. We propose (Aim 1) to define the cellular roles of PfMyoB versus PfMyoA by comparing structures, functional properties, and the role of heavy chain phosphorylation in vitro and in vivo. Aim 2 proposes to determine the binding pocket, mechanism of action, and impact on the parasite of two first-in-class small molecule inhibitors of PfMyoA ATPase activity. Aim 3 investigates two other essential Plasmodium myosins (PfMyoF and K), which are virtually unstudied both as motors and potential future druggable targets. PfMyoF likely plays a role as a processive transporter, while PfMyoK likely functions during sexual development, with motor domain inserts typical of reverse-directionality in eukaryotic class VI motors. We will use an integrative approach highlighting in vitro functional assays (motility and ensemble force assays, optical trap assays, steady- state and transient kinetics) and structural studies (X-ray crystallography and cryo-EM) together with live cell approaches (including super resolution imaging) and genetic investigation of motors (conditional knockouts/substitutions) in several stages of the Plasmodium parasite lifecycle. At completion we will have developed a previously unattainable depth of understanding into the function of the essential Plasmodium myosins as druggable targets, revealed profound insights into the structural basis by which myosins produce force and motion, and discovered fundamental insights into malaria parasite cell biology.

人类疟疾感染是由疟原虫属单细胞寄生虫引起的，是全球主要的疟疾感染之一。 健康挑战。尽管过去 15 年取得了显着进展，但全球仍有超过 4 亿人死亡 每年，其中大多数是 5 岁以下儿童。最近第一种疟疾疫苗获得许可预示着 控制疟疾的新时代，但 RTS,S 疫苗的功效相对有限意味着 如果世界卫生组织要实现到 2030 年将发病率降低 90% 的目标，那么补充方法就至关重要 意识到了。疟疾寄生虫在其复杂的人类和蚊子生命周期中都是活动的。他们移动一个 这一过程称为滑行运动，它支撑着它们到达、穿过和进入宿主组织和细胞的能力。 滑翔由寄生虫肌动球蛋白马达提供动力，破坏该马达可杀死传染性寄生虫。向 开发寄生虫肌动球蛋白运动作为可药物靶点，我们的合作团队一直致力于 表征重要的 XIV 类单头肌球蛋白马达 PfMyoA，它是滑行运动的核心。我们是 第一个表征和结晶 PfMyoA，证明其功能是由 N 端独特调节的 重链磷酸化。我们是第一个展示 PfMyoA 及其重要轻链的重要作用的人 促进红细胞 (RBC) 入侵，这是所有疟疾发病机制的原因。我们从那时起 使用 PfMyoA 突变体通过活细胞成像揭示红细胞入侵所需的能量屏障。这些 基金会专业地定位我们的团队，以扩展对整个疟疾生命周期的滑动运动的研究， 探索其他疟原虫肌球蛋白及其细胞作用，这是本次竞赛的综合目标 更新。我们建议（目标 1）通过比较结构来定义 PfMyoB 与 PfMyoA 的细胞作用， 功能特性以及重链磷酸化在体外和体内的作用。目标 2 建议 确定结合口袋、作用机制以及对两种一流小型寄生虫的影响 PfMyoA ATPase 活性的分子抑制剂。目标 3 研究另外两种重要的疟原虫肌球蛋白 （PfMyoF 和 K），它们作为马达和潜在的未来药物靶点实际上尚未被研究。肌醇 可能发挥过程转运蛋白的作用，而 PfMyoK 可能在性发育过程中发挥作用， 运动域插入是真核 VI 类运动中典型的反向性。我们将使用一个综合的 方法强调体外功能测定（运动性和整体力测定、光陷阱测定、稳态 状态和瞬态动力学）和结构研究（X 射线晶体学和冷冻电镜）以及活细胞 方法（包括超分辨率成像）和电机基因研究（条件 疟原虫寄生虫生命周期的几个阶段中的敲除/替代）。完成后我们将有 对疟原虫的功能有了前所未有的深入了解 肌球蛋白作为可药物靶标，揭示了对肌球蛋白产生的结构基础的深刻见解 力和运动，并发现了疟疾寄生虫细胞生物学的基本见解。

项目成果

Rapid tool for cell nanoarchitecture integrity assessment.

• DOI: 10.1016/j.jsb.2021.107801
• 发表时间: 2021-12
• 期刊: Journal of structural biology
• 影响因子: 3
• 作者: Gaietta G;Swift MF;Volkmann N;Hanein D
• 通讯作者: Hanein D

Mechanism of small molecule inhibition of Plasmodium falciparum myosin A informs antimalarial drug design.

• DOI: 10.1038/s41467-023-38976-7
• 发表时间: 2023-06-12
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Moussaoui, Dihia;Robblee, James P.;Robert-Paganin, Julien;Auguin, Daniel;Fisher, Fabio;Fagnant, Patricia M.;Macfarlane, Jill E.;Schaletzky, Julia;Wehri, Eddie;Mueller-Dieckmann, Christoph;Baum, Jake;Trybus, Kathleen M.;Houdusse, Anne
• 通讯作者: Houdusse, Anne

The actomyosin interface contains an evolutionary conserved core and an ancillary interface involved in specificity.

• DOI: 10.1038/s41467-021-22093-4
• 发表时间: 2021-03-25
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Robert-Paganin J;Xu XP;Swift MF;Auguin D;Robblee JP;Lu H;Fagnant PM;Krementsova EB;Trybus KM;Houdusse A;Volkmann N;Hanein D
• 通讯作者: Hanein D

Plasmodium myosin A drives parasite invasion by an atypical force generating mechanism.

疟原虫肌球蛋白 A 通过非典型的力产生机制驱动寄生虫入侵。

• DOI: 10.1038/s41467-019-11120-0
• 发表时间: 2019
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Robert-Paganin,Julien;Robblee,JamesP;Auguin,Daniel;Blake,ThomasCA;Bookwalter,CarolS;Krementsova,ElenaB;Moussaoui,Dihia;Previs,MichaelJ;Jousset,Guillaume;Baum,Jake;Trybus,KathleenM;Houdusse,Anne
• 通讯作者: Houdusse,Anne

Tunneling Nanotubes and Gap Junctions-Their Role in Long-Range Intercellular Communication during Development, Health, and Disease Conditions.

• DOI: 10.3389/fnmol.2017.00333
• 发表时间: 2017
• 期刊: Frontiers in molecular neuroscience
• 影响因子: 4.8
• 作者: Ariazi J;Benowitz A;De Biasi V;Den Boer ML;Cherqui S;Cui H;Douillet N;Eugenin EA;Favre D;Goodman S;Gousset K;Hanein D;Israel DI;Kimura S;Kirkpatrick RB;Kuhn N;Jeong C;Lou E;Mailliard R;Maio S;Okafo G;Osswald M;Pasquier J;Polak R;Pradel G;de Rooij B;Schaeffer P;Skeberdis VA;Smith IF;Tanveer A;Volkmann N;Wu Z;Zurzolo C
• 通讯作者: Zurzolo C