Mechanisms that govern assembly and function of higher order protein structures of purine metabolic enzymes

控制嘌呤代谢酶高级蛋白质结构组装和功能的机制

• 批准号: 10242969
• 负责人: Jarrod B French
• 金额: $ 38.54万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-10 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242969
• 关键词: Anabolism; Anti-Inflammatory Agents; Antimetabolites; Antiviral Agents; Biological; Biological Process; Cell Nucleus; Cell membrane; Cell physiology; Cells; Cellular Structures; Clinical; Complex; Development; Drug Targeting; Elements; Enzymes; Focal Adhesions; Foundations; Funding; Goals; Health; Human; In Vitro; Kinetics; Length; Life; Link; Metabolic; Metabolic Control; Metabolism; Microtubules; Mission; Modeling; Molecular; Multiprotein Complexes; National Institute of General Medical Sciences; Outcome Study; Output; Pathway interactions; Pharmaceutical Preparations; Physiological; Process; Production; Proteins; Purine Nucleotides; Purines; Regulation; Research; Signal Transduction; Site; Structure; System; Time; Translating; United States National Institutes of Health; anti-cancer; human disease; improved; in vivo; innovation; nucleotide metabolism; programs; protein complex; protein structure; purine metabolism; response; spatiotemporal; tool

Supramolecular complexes ranging from processing bodies to focal adhesion sites are increasingly found to be common elements of cellular structure and function. The purinosome is a recently discovered supramolecular protein complex that regulates, both temporally and spatially, the metabolism of purine nucleotides. Because of the fundamental significance of purine biosynthesis, the novelty of this type of spatiotemporal regulation, and the importance of this pathway as a drug target, there is a critical need to elucidate the mechanisms that dictate purinosome structure and function. Little is known about how such structures are formed, regulated or trafficked, nor is there a clear understanding of how these systems control metabolic flux. The long term goal of my research program is to understand how transient, supramolecular protein complexes regulate cellular metabolism. The overall objectives for the proposed funding period are to 1) define the structural features and extrinsic factors that control purinosome function, 2) quantify the kinetic and metabolic advantages that this protein structure provides, and 3) identify functional associations that purinosomes make with other cellular structures. The central hypothesis underlying these studies is that purinosome proteins undergo structural changes, in response to external signals, which drive the assembly process. In addition, we hypothesize that purinosomes are actively trafficked along microtubules, in response to specific signals, towards certain cellular structures such as the nucleus or plasma membrane. This level of control enables cells to specifically upregulate the production of purines and metabolic intermediates at a specific cellular locus. The rationale for the proposed research is that the purinosome is an important regulatory mechanism for the biosynthesis of purines, the purine biosynthetic pathway is critical to life, and is a clinically validated drug target. Thus, a better understanding of the purinosome will provide a clearer picture of overall nucleotide metabolism with potential to translate into more selective and potent antimetabolite drugs. The approach that we are taking is innovative, in the applicant’s opinion, because it departs from the status quo by integrating a suite of interdisciplinary tools to probe the system at multiple time and length scales. This will enable us to directly link molecular determinants of function with the corresponding biological outputs at physiologically relevant time and length scales. The outcome of these studies will be the elucidation of key, physiologically relevant and potentially druggable, interactions central to purinosome function, a mechanistic model that is likely generalizable to similar protein structures, and a quantitative determination of in vitro enzymatic and in vivo metabolic effects of this structure. The proposed research is significant, because it will provide sorely needed details of the structure, function and mechanism of a new paradigm in metabolic regulation – the dynamic and controllable assembly of a macromolecular protein complex of metabolic enzymes. Ultimately, the elucidation of the mechanistic underpinnings of purinosome function will help guide the development of improved anti-cancer, anti-viral and anti-inflammatory treatments.

越来越多地发现，从加工物体到焦点粘附部位的超分子络合物越来越多地发现 细胞结构和功能的常见元素。嘌呤体是最近发现的超分子 在暂时和空间上调节纯核动位剂的代谢的蛋白质复合物。由于 嘌呤生物合成的基本意义，这种类型的空间时间调节的新颖性和 该途径作为药物目标的重要性，迫切需要阐明规定的机制 嘌呤体结构和功能。关于如何形成，调节或贩运这些结构的了解知之甚少 对这些系统如何控制代谢通量，也没有清楚的了解。我的长期目标 研究计划是了解瞬时，超分子蛋白复合物如何调节细胞 代谢。拟议的资金期间的总体目标是1）定义结构特征和 控制嘌呤体功能的外在因素，2）量化这一点的动力学和代谢优势 蛋白质结构提供，3）识别嘌呤体与其他细胞产生的功能关联 结构。这些研究基础的中心假设是嘌呤体蛋白经历结构 响应外部信号的变化，这会驱动组装过程。此外，我们假设 嘌呤体被沿着微管积极地运输，以响应特定的信号，向某些细胞 核或质膜等结构。这种控制级别使单元能够专门更新 购买和代谢中间体在特定的细胞基因座的产生。提议的理由 研究是，嘌呤体是嘌呤的生物合成的重要调节机制 生物合成途径对生命至关重要，并且是临床验证的药物靶标。那，更好地理解 嘌呤体将提供更清晰的整体核苷酸代谢，并有可能转化为更多 选择性和潜在的抗代谢物药物。我们正在采用的方法是创新的，在申请人的 意见，因为它通过整合一套跨学科工具来探测系统，从而偏离了现状 在多个时间和长度尺度上。这将使我们能够直接将分子确定的功能与 在物理相关的时间和长度尺度上相应的生物输出。这些研究的结果 将是对钥匙的阐明，物理相关且潜在的吸毒的相互作用 嘌呤体功能，一种机械模型，可能可以推广到类似蛋白质结构，A 定量确定该结构的体外酶促和体内代谢作用。提议 研究很重要，因为它将提供急需的详细信息 代谢调节的新范式 - 大分子蛋白的动态和受控组装 代谢酶的复合物。最终，阐明嘌呤体的机械基础 功能将有助于指导改善抗癌，抗病毒和抗炎治疗的发展。