Regulation and Catalysis of Human Insulin Degrading Enzyme

人胰岛素降解酶的调控与催化

• 批准号: 8537935
• 负责人: WEI-JEN TANG
• 金额: $ 25.28万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8537935
• 关键词: 10q; Acetylation; Address; Adverse effects; Affect; Allosteric Regulation; Alzheimer' s Disease; Amyloid; Biochemical; Biological Assay; Biological Process; Bradykinin; Brain; CCL3 gene; CCL4 gene; Catalysis; Chemicals; Chicago; Chronic Disease; Complex; Coupled; Cysteine; Data; Development; Diabetes Mellitus; Dimerization; Drug Targeting; Endorphins; Energy Transfer; Enzyme Gene; Enzymes; Funding; Future; Genetic; Goals; Human; Human Chromosomes; Inflammation; Institutes; Insulin; Insulin-Like Growth Factor II; Insulinase; Intermediate Filament Proteins; Kinetics; Knock-out; Late Onset Alzheimer Disease; Lead; Location; Metalloproteases; Methods; Molecular; Molecular Conformation; Mus; Natriuretic Peptides; Non-Insulin-Dependent Diabetes Mellitus; Peptides; Phenotype; Phosphorylation; Physiological; Point Mutation; Post-Translational Protein Processing; Property; Proteins; Proteomics; Rattus; Regulation; Reporting; Research; Rodent; Role; Series; Single Nucleotide Polymorphism; Structure; Therapeutic; Therapeutic Agents; Transforming Growth Factors; Ubiquitin; Universities; Vimentin; Work; Yang; base; beta-Chemokines; chemokine; design; dimer; driving force; enzyme activity; enzyme mechanism; genetic analysis; human disease; hydroxamate; innovation; insight; islet amyloid polypeptide; monomer; nestin protein; non-drug; overexpression; oxidation; peptidomimetics; screening; single molecule; small molecule; success; tool

DESCRIPTION (provided by applicant): Insulin degrading enzyme (IDE) is an evolutionarily conserved, 110 kDa metalloprotease that is involved in the clearance of insulin and amyloid ? (A?). Accumulating genetic evidence in rodents and humans strongly support the role of IDE in the progression of type 2 diabetes mellitus and Alzheimer's disease. Thus, it is vital to understand the functions, catalytic mechanism, and regulation of IDE from a molecular perspective to develop viable IDE-based therapeutic strategies. We have solved the structures of human IDE in complex with insulin, A?, and other functionally relevant substrates such as natriuretic peptides and proinflammatory chemokines, CCL3/CCL4. IDE has two 55 kDa domains, which form an enclosed catalytic chamber to entrap its substrates. Our structural and biochemical analyses reveal how IDE uses an enclosed catalytic chamber to selectively recognize the global features of its substrates. The long-term goal of this research is (1) to delineate the function(s) of IDE and the mechanism(s) of its regulation and (2) to elucidate the role of IDE in human diseases. The objectives of this application are to understand (1) the molecular basis for the open-closed conformational switch and dynamics of IDE during catalysis and (2) the molecular mechanism for the regulation of IDE. In addition, we will develop potent chemical modulators of IDE to be used as potential therapeutic agents and as tools to address the biological functions of this enzyme. The central hypothesis is that the open-closed conformational switch of IDE is the key regulatory step of IDE that is subject to allosteric regulation by dimerization, posttranslational modifications, cellular factors, and chemical modulators. The rationale for the proposed research is that understanding the regulation and functions of IDE and developing small chemical modulators of IDE will ultimately allow us to better design IDE-based therapeutic strategies specific to certain human diseases such as diabetes, Alzheimer's disease, and inflammation. Guided by our preliminary data, we will study the regulation and functions of IDE in three specific aims: in Aim 1, we will use single molecule Forster resonance energy transfer analyses to address the conformational switches and dynamics that occur during catalysis of IDE and to determine how the catalysis of IDE is regulated. Aim 2 is to use two distinct screening methods to develop potent small molecule compounds that can modulate the activity of IDE and use such compounds to address the biological functions of IDE. Aim 3 is to combine structural, biochemical, and mutational studies to address the molecular basis for the regulation of IDE by dimerization, physiologically relevant cellular factors such as intermediate filament proteins, nestin and vimentin, and by posttranslational modifications such as phosphorylation and acetylation. The proposed research is significant because it will generate new insights in the dynamics and regulation of a key enzyme involved in diabetes and Alzheimer's disease and because it will also lead to the discovery of new chemical leads that can potently modulate this enzyme. The proposed research is innovative because it employs biophysical, biochemical, cellular, and medicinal chemical approaches to investigate the regulation and functions of IDE.

描述（由申请人提供）：胰岛素降解酶（IDE）是一种进化上保守的 110 kDa 金属蛋白酶，参与胰岛素和淀粉样蛋白的清除？ （一个？）。在啮齿类动物和人类中积累的遗传证据有力地支持了 IDE 在 2 型糖尿病和阿尔茨海默病进展中的作用。因此，从分子角度理解 IDE 的功能、催化机制和调控对于开发可行的基于 IDE 的治疗策略至关重要。我们已经解析了人 IDE 与胰岛素、Aβ 和其他功能相关底物（如利尿钠肽和促炎趋化因子 CCL3/CCL4）复合物的结构。 IDE 有两个 55 kDa 的结构域，形成一个封闭的催化室以捕获其底物。我们的结构和生化分析揭示了 IDE 如何使用封闭的催化室来选择性地识别其基材的整体特征。这项研究的长期目标是（1）描述 IDE 的功能及其调节机制；（2）阐明 IDE 在人类疾病中的作用。本申请的目的是了解 (1) 催化过程中 IDE 开闭构象转换和动力学的分子基础，以及 (2) IDE 调节的分子机制。此外，我们将开发 IDE 的有效化学调节剂，用作潜在的治疗剂和解决该酶生物学功能的工具。核心假设是IDE的开闭构象转换是IDE的关键调节步骤，受二聚化、翻译后修饰、细胞因子和化学调节剂的变构调节。这项研究的基本原理是，了解 IDE 的调节和功能并开发 IDE 的小型化学调节剂，最终将使我们能够更好地设计针对某些人类疾病（如糖尿病、阿尔茨海默病和炎症）的基于 IDE 的治疗策略。在我们的初步数据的指导下，我们将在三个具体目标中研究IDE的调节和功能：在目标1中，我们将使用单分子福斯特共振能量转移分析来解决IDE催化过程中发生的构象转换和动力学，并确定IDE 的催化作用是如何调控的。目标 2 是使用两种不同的筛选方法来开发可以调节 IDE 活性的有效小分子化合物，并使用此类化合物来解决 IDE 的生物学功能。目标 3 是将结构、生化和突变研究结合起来，解决通过二聚化、生理相关细胞因子（如中间丝蛋白、巢蛋白和波形蛋白）以及翻译后修饰（如磷酸化和乙酰化）调节 IDE 的分子基础。这项拟议的研究意义重大，因为它将产生关于糖尿病和阿尔茨海默氏病关键酶的动力学和调节的新见解，并且还将导致发现可以有效调节这种酶的新化学先导物。该研究具有创新性，因为它采用生物物理、生物化学、细胞和药物化学方法来研究 IDE 的调节和功能。