Project 1 - Molecular structure and function

项目1——分子结构与功能

• 批准号: 10628928
• 负责人: Francesca M Marassi
• 金额: $ 63.79万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10628928
• 关键词: Address; Affinity; Age related macular degeneration; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease patient; Amyloid beta-Protein; Atrophic; Basal lamina; Binding; Binding Proteins; Biological Assay; Blindness; Blood; Blood capillaries; Bone remodeling; Brain; Bruch' s basal membrane structure; California; Cell Adhesion; Cell Nucleus; Cell model; Cells; Chemicals; Choroid; Complement; Data; Deposition; Development; Disease; Disease Progression; Disease model; Drusen; Extracellular Matrix; Eye; Fluorescence; Glycoproteins; Goals; Hemostatic function; Hydroxyapatites; Hydroxylation; In Vitro; Investigation; Length; Link; Lipids; Medical; Minerals; Modeling; Molecular; Molecular Structure; NMR Spectroscopy; Natural Immunity; Neurons; Nuclear Magnetic Resonance; Pathologic; Process; Property; Proteins; Resolution; Retina; Risk Factors; Role; Sampling; Senile Plaques; Solid; Specificity; Structure; Structure of retinal pigment epithelium; Surface; Testing; Therapeutic Agents; Tissues; Universities; Variant; Vitronectin; biophysical techniques; calcification; calcification inhibitor; calcium phosphate; cell motility; complex biological systems; experimental study; extracellular; mineralization; novel diagnostics; novel therapeutics; reconstitution; sensor; small molecule; solid state nuclear magnetic resonance; tau Proteins; trait

PROJECT 1 SUMMARY Molecular structure and function. The ectopic accumulation of mineralized calcium-phosphate deposits in the eye and brain is a hallmark of Age-Related Macular Degeneration (AMD) and Alzheimer’s Disease (AD). In addition to inorganic components, such as hydroxyapatite (HAP), these deposits also contain lipids and proteins of both blood and intracellular origin. The process of calcification is poorly understood, and it remains unclear why some deposits progress to the disease state while others do not. It is also not clear if the associated proteins are involved in the mineralization process, or if they deposit and accumulate on the already pre-formed hydroxyapatite surface. Here, we address these important questions by comprehensive and systematic investigation of the chemical, physical and structural properties of the molecular components associated with calcified deposits, with particular focus on three validated constituents: the proteins vitronectin (Vn), amyloid-b (Ab) and Tau. Vn is a blood glycoprotein with diverse functions in hemostasis, cell adhesion and migration, innate immunity, tissue and bone remodeling. Recent studies have indicated that Vn binds both soluble Ca2+ and solid HAP with chemical specificity and the T400M Vn variant is a major risk factor for AMD. Ab and Tau are well known components of amyloid plaques in AD, but they are also found in calcification deposits in the eye and the brain. First, we will investigate the role of these proteins in the calcification process through fluorescence-based experiments and binding studies. Then, we will reconstitute proto-aggregates of HAP, lipids and proteins to model pathological deposit formation in vitro. These aggregates will provide a platform for assaying the functional roles of proteins and lipids in calcification and will aid the discovery of small molecule and biomolecular inhibitors of calcification. And finally, we will use solid-state nuclear magnetic resonance (NMR) spectroscopy to determine the structures of Vn, Tau and Ab within calcified deposits and to describe the protein-mineral interactions that drive their assembly. This information will be essential for understanding the molecular mechanism of calcification and, ultimately, for the development of new diagnostic and therapeutic agents.

项目1概要 分子结构和功能。矿化钙磷矿床的异位堆积 眼睛和大脑的损害是视网膜相关性黄斑变性（AMD）和阿尔茨海默病（AD）的标志。在 除了无机组分如羟基磷灰石（HAP）之外，这些沉积物还含有脂质， 血液和细胞内来源的蛋白质。钙化的过程知之甚少， 不清楚为什么有些沉积物进展到疾病状态，而另一些则没有。目前还不清楚， 相关的蛋白质参与矿化过程，或者如果它们存款和积累在已经 预成型的羟基磷灰石表面。在这里，我们通过全面和 系统研究分子组分的化学、物理和结构特性 与钙化沉积物相关，特别关注三种经验证的成分： (Vn)、淀粉样蛋白-b（Ab）和Tau. VN是一种血液糖蛋白，具有止血、细胞粘附、细胞增殖、细胞凋亡等多种功能 和迁移、先天免疫、组织和骨骼重塑。最近的研究表明，Vn结合了 具有化学特异性的可溶性Ca 2+和固体HAP以及T400 M Vn变体是AMD的主要危险因素。 Ab和Tau是AD中淀粉样斑块的已知组分，但它们也在钙化中发现， 沉积在眼睛和大脑里首先，我们将研究这些蛋白质在钙化过程中的作用 通过基于荧光的实验和结合研究。然后，我们将重建原始聚集体， HAP、脂质和蛋白质以模拟体外病理性存款形成。这些聚合物将提供 该平台用于测定蛋白质和脂质在钙化中的功能作用，并将有助于发现小的 钙化的分子和生物分子抑制剂。最后，我们将使用固态核磁共振 共振（NMR）光谱以确定钙化沉积物内的Vn、Tau和Ab的结构， 描述了驱动它们组装的蛋白质-矿物质相互作用。这些信息将是必不可少的， 了解钙化的分子机制，并最终开发新的诊断方法， 和治疗剂。