Structural insight into novel mechanisms of type III secretion
Structural insight into novel mechanisms of type III secretion
批准号:
10396532
负责人:
CHARALAMPOS KALODIMOS
金额:
$44.88万
依托单位国家:
美国
项目类别:
财政年份:
2011
资助国家:
美国
项目状态:
已结题
起止时间:
2011-12-01 至 2024-04-30
关键词:
ATP phosphohydrolaseAnimalsAnti-Infective AgentsBacterial ProteinsBindingBiochemicalBiological AssayBordetellaCell surfaceCellsChild MortalityComplexCryoelectron MicroscopyCytosolDataDiseaseDysenteryEventFilamentFoodFood PoisoningGram-Negative BacteriaHumanIn VitroInfantile DiarrheaKineticsLightLocomotionMembraneMolecular ChaperonesMorphologyNeedlesOrganellesPathogenesisPathogenicityPlagueProcessProtein Export PathwayProtein SecretionProteinsProton-Motive ForcePseudomonasRecyclingRegulationReportingResolutionRoleSalmonellaShigellaSignal TransductionSorting - Cell MovementSpecificityStructureSystemThermodynamicsTimeType III Secretion System PathwayTyphoid FeverVaccinesVirulenceVirulence FactorsX-Ray Crystallographycell motilitydesignenteropathogenic Escherichia coliin vivoinsightkinetosomenanomachinenoveloperationpathogenic Escherichia colipathogenic bacteriaprotein structurestructural biologytherapeutic protein
中文摘要
项目摘要
III型分泌系统(T3SS)是一种纳米机器,专门用于革兰氏阴性细菌的蛋白质输出。
T3SS具有相同的形态和整体结构,在功能上可以分为两个进化相关的fi
分类:flagella T3SS,它促进细菌的运动和由flagellum实现的运动,以及致病
T3SS,它使用注射体将毒力蛋白输送到人类或动物宿主细胞中。
在过去的十年里,在理解fi的结构、组装和模式方面取得了显著的进展
T3SS的操作。fl凝胶体和注射体的主要结构构建蛋白
嵌入到细胞表面突出的fi尖端的内外细菌膜中,以及
细胞液成分已经被广泛地表征。鞭毛蛋白和毒力因子(效应器、针头
蛋白质和转运体)在细胞质中与T3S专属的伴侣形成紧密的络合物,随后
靶向fi定位于位于膜上的出口装置。在ATP和质子推动力的推动下,
然后,fl琼脂蛋白和细菌效应器通过该通道转运。关于以下方面的基本问题
支撑这些进程的功能机制仍未得到解决。
我们建议使用结构、动力学、热力学、动力学、生化和生物化学相结合的综合方法
体外和体内的功能分析,以提供对易位过程的早期事件的洞察,涉及
分子伴侣识别机制,针对ATPase和分选平台的靶向机制,选择
控制fi撕裂形成蛋白和效应器的分层运输的机制,最终
组装机械的胞液部分。在过去的几年里,我们对T3S蛋白进行了广泛的表征
肠道致病性大肠埃希氏菌(EPEC)的成分,婴儿腹泻和儿童死亡的主要原因
世界范围内,以及沙门氏菌属。通常与食物中毒有关。我们在这里提出了一种新的fi代码,它支持
关于T3SS执行其功能的机制的非常耐人寻味的假设。fic++的具体目标是
旨在提供对(I)关键t3之间以及关键t3之间的specific相互作用的机制的原子分辨率洞察
蛋白质,(2)将T3S蛋白质靶向出口门的机制基础,(3)“识别”和“分泌”
(4)出口门的装配和操作机构,最终是整个T3S机械的装配和操作机构。
英文摘要
Project Summary
Type III secretion systems (T3SSs) are nanomachines that are dedicated to protein export in Gram-negative bacteria.
T3SSs share the same morphology and overall structure and can be functionally classified into two evolutionary-related
classes: the flagellar T3SS, which promotes bacterial locomotion and motility enabled by the flagellum, and the pathogenic
T3SS, which uses the injectisome to transport virulence proteins into human or animal host cells.
Over the past decade significant progress has been made in understanding the structure, assembly and the mode of
operation of T3SS. The principal structural building proteins of the flagellum and the injectisome, from the basal body
embedded in the inner and outer bacterial membrane to the tip of the filaments protruding from the cell surface, and the
cytosolic components have been extensively characterized. Flagellar proteins and virulence factors (effectors, needle
proteins and translocators) form tight complexes with T3S-dedicated chaperones in the cytosol and are subsequently
targeted specifically to the export apparatus located at the membrane. Powered by ATP and the proton motive force, the
flagellar proteins and bacterial effectors are then translocated through the channel. Fundamental questions about the
functional mechanisms underpinning these processes remain unaddressed.
We propose to use an integrated approach combining structural, dynamic, thermodynamic, kinetic, biochemical and in
vitro and in vivo functional assays to provide insight into the early events of the translocation process that involve the
recognition mechanisms by chaperones, targeting mechanisms to the ATPase and the sorting platform, selection
mechanisms that control the hierarchical transport of the filament-forming proteins and the effectors and ultimately the
assembly of the cytosolic part of the machinery. We have extensively characterized over the last years T3S protein
components from the enteropathogenic Escherichia coli (EPEC), the major cause of infantile diarrhea and child mortality
worldwide, as well as from Salmonella sp. commonly associated with food poising. We present here novel findings supporting
very intriguing hypotheses about the mechanisms used by T3SSs to carry out their function. The specific aims are
designed to provide atomic-resolution insight into (i) the mechanisms of specific interactions between and among key T3S
proteins, (ii) the mechanistic basis for targeting of T3S proteins to the export gate, (iii) the “recognition” and “secretion”
signal, and (iv) the assembly and operation mechanisms of the export gate and ultimately of the entire T3S machinery.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Structure and functional mechanisms of molecular chaperones and protein kinases
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批准号:9920188
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资助金额:$75.38万
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Allosteric and Transport Mechanisms in TonB-dependent Transporters
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Structural insight into novel mechanisms of type III secretion
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Structural insight into novel mechanisms of type III secretion
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Structural insight into novel mechanisms of type III secretion
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Structural insight into novel mechanisms of type III secretion
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Structural insight into novel mechanisms of type III secretion
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批准号:10609868
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资助金额:$44.88万
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财政年份:2011
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Regulatory and recognition mechanisms of translocase ATPase SecA
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资助金额:$11.55万
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财政年份:2010
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依托单位:
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财政年份:2009
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依托单位:
STRUCTURAL STUDIES OF THE 70 KDA SECB CHAPERONE
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依托单位:
STRUCTURAL STUDIES OF THE 70 KDA SECB CHAPERONE
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资助金额:$0.4万
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财政年份:2008
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依托单位:
Regulatory and recognition mechanisms of translocase ATPase SecA
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海外基金