Complex Dynamics in Multisite Phosphorylation

多位点磷酸化的复杂动力学

• 批准号: 7614488
• 负责人: JEREMY GUNAWARDENA
• 金额: $ 35.13万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-15 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614488
• 关键词: Behavior; Biochemical; Biochemistry; Biological Models; Cell physiology; Clinical; Complex; Development; Enzymes; Equilibrium; Exhibits; Goals; Malignant Neoplasms; Mass Spectrum Analysis; Modeling; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Population; Protein Dephosphorylation; Protocols documentation; Role; System; Testing; human disease; insight; interdisciplinary approach; multidisciplinary; novel; public health relevance

DESCRIPTION (provided by applicant): Multisite protein phosphorylation and dephosphorylation (P&D) are fundamental to most eukaryotic cellular processes. Disregulation of multisite P&D is implicated in several human diseases, including cancer, and both kinases and phosphatases are targets of important drugs in clinical use or development. The goal of this proposal is to investigate the dynamical complexity that arises when kinases and phosphatases collectively regulate the cellular phosphorylation state. A substrate molecule with n phosphorylation sites may occupy 2n states and a population of substrate molecules will have a distribution of such phospho-forms, with the system of kinase, phosphatase and substrate being maintained away from equilibrium by a steady supply of ATP. We will develop a mass spectrometry protocol to distinguish all 2n phospho-forms and assemble a set of model P&D systems, having varying numbers of phosphorylation sites, enzymatic mechanisms and cellular roles, on which to undertake systems biochemistry. We will determine mathematically the conditions under which such systems exhibit complex behaviors, such as multistability and oscillations, and will test these predictions on the model systems. Such a multidisciplinary approach will provide novel insights into these complex, medically important and poorly-understood systems. PUBLIC HEALTH RELEVANCE: Disregulation of protein phosphorylation and dephosphorylation is implicated in several human diseases, including cancer, and the enzymes involved, kinases and phosphatases, are targets of important drugs in clinical use and development. Systems of kinase, phosphatase and substrate show surprising complexity, especially as the number of phosphorylation sites increases. This proposal seeks to investigate such complexity through a multidisciplinary mathematical and experimental approach, using several model phosphorylation and dephosphorylation systems having varying cellular roles, enzymatic mechanisms and numbers of phosphorylation sites.

描述（由申请人提供）：多位点蛋白质磷酸化和去磷酸化（P&D）是大多数真核细胞过程的基础。多位点 P&D 失调与包括癌症在内的多种人类疾病有关，激酶和磷酸酶都是临床使用或开发中重要药物的靶点。该提案的目标是研究激酶和磷酸酶共同调节细胞磷酸化状态时产生的动态复杂性。具有 n 个磷酸化位点的底物分子可以占据 2n 个状态，并且底物分子群体将具有此类磷酸化形式的分布，其中激酶、磷酸酶和底物的系统通过稳定的 ATP 供应而保持远离平衡。我们将开发一种质谱方案来区分所有 2n 磷酸形式并组装一组模型 P&D 系统，该系统具有不同数量的磷酸化位点、酶促机制和细胞作用，并在其上进行系统生物化学。我们将从数学上确定此类系统表现出复杂行为（例如多稳定性和振荡）的条件，并将在模型系统上测试这些预测。这种多学科方法将为这些复杂的、具有医学重要性且人们知之甚少的系统提供新颖的见解。公共健康相关性：蛋白质磷酸化和去磷酸化的失调与包括癌症在内的多种人类疾病有关，而所涉及的酶、激酶和磷酸酶是临床使用和开发中重要药物的靶标。激酶、磷酸酶和底物系统显示出令人惊讶的复杂性，特别是随着磷酸化位点数量的增加。该提案旨在通过多学科数学和实验方法，使用具有不同细胞作用、酶机制和磷酸化位点数量的几种模型磷酸化和去磷酸化系统来研究这种复杂性。