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 摘要 矿化磷酸钙沉积物的分子结构和功能。 眼睛和大脑是年龄相关性黄斑变性 (AMD) 和阿尔茨海默病 (AD) 的标志。 除了羟基磷灰石 (HAP) 等无机成分外，这些沉积物还含有脂质和 血液和细胞内来源的蛋白质对钙化的过程知之甚少，而且这一过程仍然存在。 目前还不清楚为什么有些沉积物会发展为疾病状态，而另一些则不会。 相关蛋白质参与矿化过程，或者如果它们沉积并积累在已经 在这里，我们通过综合和解决这些重要问题。 系统研究分子成分的化学、物理和结构特性 与钙化沉积物相关，特别关注三种经过验证的成分：玻连蛋白 (Vn)、淀粉样蛋白-b (Ab) 和 Tau 是一种血液糖蛋白，具有止血、细胞粘附等多种功能。 最近的研究表明，Vn 与两者结合。 可溶性 Ca2+ 和具有化学特异性的固体 HAP 以及 T400M Vn 变体是 AMD 的主要危险因素。 Ab 和 Tau 是 AD 中淀粉样斑块的众所周知的成分，但它们也存在于钙化中 首先，我们将研究这些蛋白质在钙化过程中的作用。 然后，我们将通过基于荧光的实验和结合研究来重建原聚集体。 HAP、脂质和蛋白质可模拟体外病理沉积物的形成。 用于测定蛋白质和脂质在钙化中的功能作用的平台，并将有助于发现小的 最后，我们将使用固态核磁。 核磁共振 (NMR) 光谱法可确定钙化沉积物中 Vn、Tau 和 Ab 的结构，并 描述驱动其组装的蛋白质-矿物质相互作用。 了解钙化的分子机制，并最终开发新的诊断方法 和治疗剂。