ENDOTHELIAL ALPHA V INTEGRIN COMPLEXES MODULATED ACTIN CYTOSKELETAL ORGANIZATION

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 整联蛋白是细胞表面受体，具有许多基本的细胞功能。 这些功能之一，这是相关的器官水肿，发生在炎症性疾病，如急性肺损伤，是调节流体和溶质通过内皮细胞，细胞层内衬血管在整个循环系统。 Sheppard实验室最近证明，α-v整合素通过组织肌动蛋白细胞骨架来调节内皮通透性。 具体而言，α-v β-3整联蛋白是肌动蛋白重排到皮质分布所必需的，其响应于鞘氨醇1-磷酸（一种血浆携带的屏障增强分子）而增加血管屏障强度;另一方面，α-v β-5是细胞骨架组织成应力纤维所必需的，其响应于凝血酶而降低屏障强度，凝血酶是增加血管渗漏的血液携带的介质。 我们的目标是确定与这些整合素相关的蛋白质复合物的成分。 在这方面，一种理想的方法是采用质谱法，其中可以将生化分离的整合素复合物提交给质谱测序以用于蛋白质发现。 与这种公正的方法，不同的肌动蛋白结合和/或调节蛋白与这两个整合素可以被确定和进一步研究，以了解他们如何调节肌动蛋白的组织。 有了这些强有力的信息，我们可以进一步剖析介导这两种整合素的相反生物反应的途径，并沿着其各自的途径靶向特定点，作为脓毒症和其他急性炎症性疾病背景下器官水肿的潜在治疗方法。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Integrins are cell surface receptors that serve a number of basic cellular functions. One of these functions, which is relevant to the organ edema that occurs in inflammatory diseases like acute lung injury, is the regulation of passage of fluid and solute across the endothelium, the layer of cells lining blood vessels throughout the circulatory system. The Sheppard lab has recently demonstrated that alpha-v integrins modulate endothelial permeability by organizing the actin cytoskeleton. Specifically, alpha-v beta-3 integrin is necessary for rearrangement of actin into cortical distribution that increases vascular barrier strength in response to sphingosine 1-phosphate, a plasma borne barrier-enhancing molecule; on the other hand, alpha-v beta-5 is required for organization of the cytoskeleton into stress fibers that decrease barrier strength in response to thrombin, a blood borne mediator of increased blood vessel leakiness. Our objective is to identify the constituents of the protein complexes associated with these integrins. An ideal approach in this regard is to employ mass spectrometry, where biochemically isolated integrin complexes can be submitted to mass spectrometric sequencing for protein discovery. With this unbiased approach, the distinct actin binding and/or modulatory proteins associated with these two integrins may be identified and studied further to understand how they modulate actin organization. Armed with this powerful information, we may further dissect pathways mediating the opposing biological response of these two integrins and target specific points along their respective pathways as potential treatments for organ edema in the setting of sepsis and other acute inflammatory diseases.