AFAP-110 AS A REGULATOR OF ANGIOGENESIS

AFAP-110 作为血管生成的调节剂

• 批准号: 7610250
• 负责人: ROBERT SHURINA
• 金额: $ 19.67万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610250
• 关键词: Adaptor Signaling Protein; Adhesions; Affect; Behavior; Binding; Blood Vessels; COS Cells; Cell Shape; Cell membrane; Cells; Chimeric Proteins; Computer Retrieval of Information on Scientific Projects Database; Cytoskeleton; Dominant-Negative Mutation; Endothelial Cells; Equus caballus; Event; Exhibits; Funding; Grant; Green Fluorescent Proteins; In Vitro; Institution; Link; Measures; Microfilaments; Mutation; Pathway interactions; Protein Binding; Protein Kinase C; Proteins; Rattus; Recombinant Fusion Proteins; Recombinant Proteins; Research; Research Personnel; Resources; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Source; United States National Institutes of Health; Vascular Endothelial Growth Factors; angiogenesis; cell motility; prevent; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. AFAP-110 as a regulator of angiogenesis Cell signaling in angiogenesis involves an interplay of signaling pathways that culminate in the formation of new blood vessels. Cell migration, adhesion and microvessel formation are necessary events in angiogenesis, and involve changes in a cell?s shape that occur through alterations in the cytoskeleton. Src and PKC affect changes in actin filaments by interacting with the actin filament associated protein (AFAP). Both Src and protein kinase C (PKC) activation are important in vascular endothelial growth factor (VGEF) pathways that stimulate angiogenesis, although their roles are poorly understood. AFAP is a 110-kilodalton-adaptor protein that links the signaling molecules PKC and Src to actin filaments. Our group has shown that AFAP binds to cSrc and activates it. Deletions in AFAP enable it to activate cSrc. Also, PKC activators direct changes in AFAP that enable it to activate cSrc. Mutations that block interactions with cSrc or interactions with PKC prevent AFAP from activating cSrc in response to PKC?. Since AFAP is expressed at high levels in endothelial cells, we hypothesize a role for AFAP in angiogenesis. We assess its role in angiogenesis by creating cell permeable fusion proteins for the wild type, dominant negative and dominant positive forms of AFAP. Fusion proteins will be constructed by inserting an Antennepedia sequence in frame with GFP-AFAP. The green fluorescent protein (GFP) will allow us to demonstrate that the target cell has taken up the recombinant protein. We have previously shown that the GFP-AFAP fusion protein is functional in COS cells and exhibits identical behavior to AFAP regarding PKC binding and Src activation. Antennapedia sequences have been shown to serve as ?Trojan horses? that can deliver cargo proteins across cell membranes. The ability of the recombinant fusion proteins to stimulate angiogenesis in vitro will be measured using endothelial cells or rat aortic ring explant cultures.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 AFAP-110作为血管生成的调节剂 血管生成中的细胞信号传导涉及信号传导途径的相互作用，最终导致新血管的形成。 细胞迁移、粘附和微血管形成是血管生成过程中必不可少的事件，并涉及细胞内的变化。的形状，通过改变细胞骨架发生。 Src和PKC通过与肌动蛋白丝相关蛋白（AFAP）相互作用影响肌动蛋白丝的变化。 Src和蛋白激酶C（PKC）激活在刺激血管生成的血管内皮生长因子（VGEF）通路中很重要，尽管它们的作用知之甚少。 AFAP是一种110千道尔顿的衔接蛋白，它将信号分子PKC和Src连接到肌动蛋白丝上。 我们的研究小组已经证明AFAP与cSrc结合并激活它。 AFAP中的删除使其能够激活cSrc。 此外，PKC激活剂指导AFAP的变化，使其能够激活cSrc。 阻断与cSrc相互作用或与PKC相互作用的突变阻止AFAP激活cSrc以响应PKC？。 由于AFAP在内皮细胞中高水平表达，我们推测AFAP在血管生成中的作用。 我们通过创建野生型、显性阴性和显性阳性形式的AFAP的细胞可渗透融合蛋白来评估其在血管生成中的作用。 融合蛋白将通过在具有GFP-AFAP的框中插入一个噬菌体序列来构建。 绿色荧光蛋白（GFP）将使我们能够证明靶细胞已经吸收了重组蛋白。 我们先前已经证明，GFP-AFAP融合蛋白在COS细胞中是功能性的，并且在PKC结合和Src激活方面表现出与AFAP相同的行为。 已经证明了昆虫足的序列可以作为？特洛伊木马？可以将货物蛋白质运送到细胞膜上。 将使用内皮细胞或大鼠主动脉环外植体培养物测量重组融合蛋白刺激体外血管生成的能力。