权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Structural characterization of OM proteins from Gram-negative pathogens

革兰氏阴性病原体 OM 蛋白的结构表征

基本信息

批准号：
8741336
负责人：
Susan Buchanan
金额：
$ 61.73万
依托单位：
NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8741336
关键词：
Adverse effects Air Amendment Animals Antibiotic Resistance Antibiotic Therapy Antibiotics Antigens Attention Bacteria Bacteriophage T4 Bacteriophages Biological Biology Bite Bubonic Plague Cells Cessation of life Communicable Diseases Development Disease Disease Outbreaks Drug Targeting Drug resistance Economics Effectiveness Engineering Epidemic European Fleas Human Hybrids Hygiene In Vitro Individual Iron Knowledge Lung Lysine Membrane Membrane Proteins Microbiology Monitor Muramidase Names Nature Patients Peptidoglycan Pesticin Pharmaceutical Preparations Pharmacotherapy Plague Plague Vaccine Pneumonic Plague Preventive Production Proteins Publishing Recording of previous events Recurrence Reporting Research Resistance Rodent Siderophores Speed Structure System Testing Time Toxin Travel Update Vaccination Vaccines Virulence Virulent Whole Cell Vaccine Work World Health Organization Yersinia Yersinia pestis bacteriocin base capsule cell killing contagion improved in vivo innovation interest killings lysin middle age mouse model novel novel vaccines pathogen periplasm polypeptide programs resistant strain respiratory vaccine development weapons yersiniabactin

项目摘要

Our structures of Y. pestis FyuA and pesticin allowed us to engineer a novel phage therapy drug against a Gram-negative pathogen: We solved the structure of FyuA, a TonB-dependent iron transporter required for virulence in bubonic plague, with and without its cognate siderophore, ferric yersiniabactin. At the same time, we determined the structure of a bacteriocin called pesticin that uses FyuA to cross the outer membrane. Once inside the periplasm, pesticin kills the cell by degrading the peptidoglycan layer. From our structure we discovered that the killing domain of pesticin resembles phage T4 lysozyme, so we engineered a hybrid bacterial-phage toxin that contains a bacterial targeting domain (to FyuA) and a phage killing domain. We showed that the hybrid lysine evades the natural protection mechanism of toxin-producing strains and kills all Yersinia strains tested, both in vitro and in vivo (mouse model of bubonic plague). This is the first demonstration of phage therapy for Gram-negative pathogens because until now, no one knew how to transport the toxin across the outer membrane. This work was recently published at PNAS* and has received considerable attention, including a Nature Microbiology Reviews Highlight. We have recently begun a project to identify and characterize outer membrane proteins that are either essential or required for virulence in a variety of Gram-negative pathogens, since these proteins may make good vaccine components and drug targets. Structural characterization of several candidate proteins is currently underway. *Lukacik, P., Barnard, T.J., Keller, P.W., Chaturvedi, K., Seddiki, N., Fairman, J.W., Noinaj, N., Kirby, T.L., Henderson, J.P., Steven, A.C., Hinnebusch, B.J. & Buchanan, S.K. (2012). Structural engineering of a phage lysin that targets Gram-negative pathogens. Proc. Natl. Acad. Sci. USA, 109, 9857-9862. PMCID: PMC3382549 Research Highlighted: In this Issue Proc. Natl. Acad. Sci. USA, 109, 9667-9668 (2012). Research Highlighted: Nat. Rev. Microbiol. 10, 520-521 (2012). Research Highlighted: SciBX 5(27); doi:10.1038/scibx.2012.706 (2012) (Nature publishing group).

Y.鼠疫FyuA和pesticin使我们能够设计一种针对革兰氏阴性病原体的新型噬菌体治疗药物：我们解决了FyuA的结构，一个TonB依赖的铁转运蛋白所需的毒力在腺鼠疫，有和没有其同源铁载体，铁耶尔森菌素。与此同时，我们确定了一种名为pesticin的细菌素的结构，它使用FyuA穿过外膜。一旦进入周质，pesticin通过降解肽聚糖层杀死细胞。从我们的结构中，我们发现pesticin的杀伤结构域类似于噬菌体T4溶菌酶，因此我们工程化了含有细菌靶向结构域（针对FyuA）和噬菌体杀伤结构域的杂合细菌-噬菌体毒素。我们表明，杂合赖氨酸逃避天然保护机制的产毒菌株和杀死所有耶尔森氏菌菌株测试，在体外和体内（小鼠模型的腺鼠疫）。这是第一次证明噬菌体治疗革兰氏阴性病原体，因为到目前为止，没有人知道如何将毒素运输穿过外膜。这项工作最近发表在PNAS* 上，受到了相当大的关注，包括Nature Microbiology Reviews Highlight。我们最近开始了一个项目，以确定和表征外膜蛋白，这些蛋白是必不可少的或需要在各种革兰氏阴性病原体的毒力，因为这些蛋白质可能成为良好的疫苗成分和药物靶点。目前正在对几种候选蛋白质进行结构表征。 *Lukacik，P.，Barnard，T.J.，凯勒，P.W.，Chaturvedi，K.，Seddiki，N.，费尔曼，J.W.，Noinaj，N.，Kirby，T.L.，亨德森，J. P.，史蒂文，AC，Hinnebusch，B.J. & Buchanan，S.K.（2012年）。革兰氏阴性病原体靶向噬菌体溶素的结构工程。Proc. Natl. Acad. Sci. USA，109，9857-9862. PMCID：PMC3382549 研究突出显示：在这个问题上。Acad. Sci. USA，109，9667-9668（2012）。研究重点：自然微生物学。10，520-521（2012）中所述。 Research Highlighted：SciBX 5（27）; doi：10.1038/scibx.2012.706（2012）（Nature publishing group）.