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.