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.

由疟原虫属单细胞寄生虫引起的人类疟疾感染是全球性的重大疾病

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