Protein Interactions and Conformational Changes in Fibronectin Fibril Formation

纤连蛋白原纤维形成中的蛋白质相互作用和构象变化

• 批准号: 9134853
• 负责人: Klara Briknarova
• 金额: $ 28.41万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9134853
• 关键词: Affinity; Age related macular degeneration; Angiogenesis Inhibition; Angiogenesis Inhibitors; Architecture; Atherosclerosis; Binding; Binding Sites; Biological Models; Blood Vessels; Cell Adhesion; Cells; Chronic; Complex; Cysteine; Data; Deposition; Development; Diabetic Retinopathy; Disease; Disulfides; Energy Transfer; Extracellular Matrix; Extracellular Matrix Proteins; Fibronectins; Generations; Goals; Growth; Health; In Vitro; Investigation; Label; Lesion; Light; Malignant Neoplasms; Maps; Microscopic; Molecular; Molecular Conformation; Molecular Models; NMR Spectroscopy; Neoplasm Metastasis; Pathologic Neovascularization; Pathologic Processes; Physiological Processes; Play; Process; Protein Dynamics; Proteins; Proteolysis; Recruitment Activity; Regulation; Relaxation; Reproduction; Resolution; Rheumatoid Arthritis; Role; Signal Transduction; Site; Site-Directed Mutagenesis; Structural Genes; Structure; Vascular blood supply; Wound Healing; X-Ray Crystallography; angiogenesis; base; beta pleated sheet; crosslink; fibrillogenesis; in vivo; inhibitor/antagonist; insight; migration; molecular modeling; molecular rearrangement; mouse model; mutant; neoplastic cell; new growth; novel anticancer drug; single molecule; tumor; tumor growth; tumor vascular supply

 DESCRIPTION (provided by applicant): The generation of a lethal tumor mass requires that the tumor cells recruit and sustain their own blood supply. In the absence of blood supply, tumors can persist as dormant microscopic lesions but they do not expand beyond the size of 1-2 mm. Hence, inhibition of new blood vessel growth (angiogenesis) offers great promise to eradicate tumors or to convert cancer to a chronic manageable disease. In addition, inhibition of angiogenesis offers great promise to treat a range of other diseases that depend on angiogenesis, including age-related macular degeneration, diabetic retinopathy, rheumatoid arthritis and atherosclerosis. The focus of this application is anastellin, a fragment of the extracellular matrix protein fibronectin that inhibits angiogenesis, tumor growth and metastasis in mouse models. Anastellin requires endogenous fibronectin for its in vivo anti- angiogenic activity, and it binds to fibronectin in vitro and converts the soluble protein to insoluble fibril. Our long term goal is to elucidate how the interaction of anastellin with fibronectin leads to formation of fibrillar aggregates, and in general how the structure of fibronectin determines its function. The goal of this application is to identify the molecular basis of the interaction betwee anastellin and fibronectin. We propose to investigate the structure and dynamics of the complexes between anastellin and its target fibronectin type III (FN3) domains by disulfide crosslinking, nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography. In addition, we propose to investigate the conformation of the anastellin binding region in fibronectin using paramagnetic relaxation enhancement (PRE) and single molecule Förster resonance energy transfer (FRET). Identification of structural features that are responsible for the activity of anastellin will provide detailed understanding of how this inhibitor of angiogenesi acts at the molecular level and may enable development of new anticancer drugs. The proposed structural studies will also provide insight into the molecular interactions and rearrangements that are involved in conversion of soluble fibronectin to the fibrillar matrix form.

 描述（由适用提供）：致死性肿瘤质量的产生要求肿瘤细胞募集和维持自己的血液供应。在没有血液供应的情况下，肿瘤可以持续为休眠的微观病变，但不会膨胀超过1-2 mm。因此，抑制新血管生长（血管生成）为放射性肿瘤或将癌症转化为慢性可管理疾病提供了巨大的希望。此外，抑制异性生物质的抑制作用提供了巨大的希望，可以治疗依赖血管生成的一系列其他疾病，包括与年龄相关的黄斑变性，糖尿病性视网膜病，类风湿关节炎和动脉粥样硬化。该应用的重点是anastellin，这是细胞外基质蛋白纤连蛋白的片段，可抑制小鼠模型中的血管生成，肿瘤生长和转移。阿纳斯特蛋白需要内源性纤连蛋白的体内抗血管生成活性，并在体外与纤连蛋白结合，并将固体蛋白转化为不溶性原纤维。我们的长期目标是阐明阿纳斯特蛋白与纤连蛋白的相互作用如何导致形成原纤维聚集体的形成，并且通常如何确定纤连蛋白的结构如何确定其功能。该应用的目的是确定anastellin和纤连蛋白之间相互作用的分子基础。我们建议通过二硫键交联，核磁共振（NMR）光谱和X射线晶体学来研究Anastellin及其靶纤维蛋白III型（FN3）型型结构域之间复合物的结构和动力学。此外，我们建议使用顺磁性弛豫增强（PRE）和单分子försterresonance能量转移（FRET）研究纤连蛋白中anastellin结合区域的构象。鉴定负责阿纳斯特蛋白活性的结构特征将详细了解这种血管生成剂在分子水平上如何起作用，并可能能够开发新的抗癌药物。提出的结构研究还将提供有关固体纤连蛋白转化为原纤维基质形式的分子相互作用和重排的洞察力。