Direct Rap1-talin interaction in platelets, leukocytes, and endothelial cells
Direct Rap1-talin interaction in platelets, leukocytes, and endothelial cells
批准号:
10229368
负责人:
Mark HOWARD Ginsberg
金额:
$48.79万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-08-05 至 2025-07-31
关键词:
AchievementAdhesionsAgeAgonistAllelesBindingBinding SitesBiological AssayBloodBlood CellsBlood PlateletsBlood VesselsCarotid ArteriesCell AdhesionCell physiologyCellsCollaborationsDataDevelopmentDisabled PersonsEmbryoEndothelial CellsEndotheliumFoundationsHemorrhageHemostatic functionHomingIn VitroIndividualInflammationIntegrinsIntercellular JunctionsKnockout MiceLaser injuryLeukocytesLigandsLungLymphoid TissueMediatingMegakaryocytesModelingMolecular ConformationMusMutationNaturePathway interactionsPeripheralPhenotypePlayRegulatory T-LymphocyteRoleSignal TransductionSiteStenosisStructureT-LymphocyteTalinTamoxifenTestingThrombosisTumor AngiogenesisVena caval structureVenous Thrombosisbrain endothelial cellcell typeferric chloridein vivomutantneutrophiloverexpressionplatelet functionrecruit
中文摘要
摘要
Rap1是一个连接激动剂刺激和Talin募集到整合素的中枢信号节点,a
整合素激活的最后一个共同步骤。在白细胞中,RIAM是一种RAP1效应器,它介导
Talin依赖的激活;然而,在血小板中,这种Rap1效应器的身份尚不清楚
这是我们理解上的一个关键差距。初步数据表明,一种新的进化保守的
该范式认为talin包含两个Rap1结合位点,使其能够作为Rap1效应器。这个
申请人假设Talin本身是牵连的主要且可能是唯一的Rap1效应器
在血小板整合素的激活中。其次,他认为Talin-Rap1的相互作用可能
有助于整合素功能,即使在含有丰富的RAP1效应器的白细胞中也是如此
而在含有大量片层脂蛋白的内皮细胞中,这是一种RIAM类似物。检视
这些想法:特定目标1将检验直接的Rap1-talin相互作用发挥作用的假设
在血小板整合素活化中起核心作用。小鼠的巨核细胞和血小板
具有任一个或两个RAP1结合位点被禁用的表达TALIN将被分析血小板结构,
功能和编队。止血和血栓形成将通过各种化验进行测试。一个
生物膜力探针将表征Rap1-talin相互作用在个体中的作用
IIb3整合素-配位键。《特定目标2》将检验一种假说,即引导Rap1-Talin
相互作用在充满RIAM的细胞中很重要。与项目2整合协作
白细胞的激活、构象和拓扑结构将在携带每一种
Aim1中描述的三个Talin等位基因。每种突变与RIAM联合应用的效果
缺失或RIAM过度表达将测试这两个RAP1效应器的相对作用。
特定目标3将检验Talin-Rap1相互作用在
内皮细胞。选择性表达三种Talin突变等位基因之一的小鼠
将研究血管内皮细胞的出血和发育表型。对整合素的影响
将在体外评估原发肺和脑的激活、扩散和细胞-细胞连接。
微血管内皮细胞。LamelLipodin的缺失和过度表达将使分析成为可能
内皮细胞功能中层脂蛋白和Rap1结合与talin的关系。一起,
这些目标的实现将检验塔林本身作为范式转换假说
血小板功能中RAP1的主要效应分子在止血和血栓形成中的作用
Rap1-talin相互作用在白细胞和内皮细胞中的重要性。
英文摘要
Abstract
Rap1 is a central signaling node connecting agonist stimulation to talin recruitment to integrins, a
final common step in integrin activation. In leukocytes, RIAM is a Rap1 effector that mediates
talin-dependent activation; however, in platelets the identity of such Rap1 effectors is obscure
and is a key gap in our understanding. Preliminary data suggest a new evolutionarily-conserved
paradigm that talin contains two Rap1 binding sites that enable it to serve as a Rap1 effector. The
applicant hypothesizes that talin itself is the principal, and perhaps only, Rap1 effector implicated
in platelet integrin activation. Secondly, he suggests that that the talin-Rap1 interaction may
contribute to integrin function even in leukocytes that contain an abundant Rap1 effector, RIAM
and in endothelial cells that contain substantial lamellipodin, a RIAM paralogue. To examine
these ideas: Specific Aim 1 will test the hypothesis that a direct Rap1-talin interaction plays
a central role in platelet integrin activation. Mice in which the megakaryocytes and platelets
express talin with either or both Rap1 binding sites disabled will be analyzed for platelet structure,
function, and formation. Hemostasis and thrombosis will be tested in a variety of assays. A
biomembrane force probe will characterize the role of the Rap1-talin interaction in individual
IIb3 integrin-ligand bonds. Specific Aim 2 will test the hypothesis that direct Rap1-Talin
interaction is important in RIAM-replete cells. In collaboration with Project 2 Integrin
activation, conformation, and topology in leukocytes will be analyzed in cells bearing each of the
three talin alleles described in Aim1. The effect of each mutation in combination with RIAM
deletion or with RIAM over-expression will test the relative roles of these two Rap1 effectors.
Specific Aim 3 will test the hypothesis that the talin-Rap1 interaction is important in
endothelial cells. Mice expressing one of the three mutant talin alleles selectively in
endothelium will be studied for hemorrhage and developmental phenotypes. Effects on integrin
activation, spreading, and cell-cell junctions will be assessed in vitro in primary lung and brain
microvascular endothelial cells. Lamellipodin deletion and over expression will enable an analysis
of the relationship of lamellipodin and Rap1 binding to talin in endothelial cell functions. Together,
achievement of these aims will test the paradigm-shifting hypothesis that talin itself serves as
the major Rap1 effector in platelet function in hemostasis and thrombosis and evaluate the
importance of the Rap1-talin interaction in leukocytes and endothelial cells.
期刊论文(0)
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科研奖励(0)
会议论文
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Anti-Coagulant and Cytoprotective activity in CCM pathogenesis
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Anti-Coagulant and Cytoprotective activity in CCM pathogenesis
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Composition and Functions of the Integrin Activation Complex
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海外基金