EAGER: Nature of the Pre-chemistry Ensemble in Protein Kinases

EAGER：蛋白激酶中前化学整体的本质

• 批准号: 1811770
• 负责人: Ranajeet Ghose
• 金额: $ 29.95万
• 依托单位: CUNY City College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1811770&HistoricalAwards=false
• 关键词: EAGER; Nature; Pre; chemistry; Ensemble

Kinases are biological molecules that modify the rate of specific chemical reactions in cells. They and the products of these chemical reactions represent central hubs in the intracellular signaling networks in all domains of life. In addition to sub-cellular localization and temporal expression, the efficiencies of particular kinases towards specific molecular products is key in targeting input signals towards distinct physiological outputs. The goal of this project is to understand the nature of this regulation with sufficient resolution in space and time to provide the means to perturb and ultimately design signaling networks that drive specific input signals towards defined cellular responses. This project will also provide a fertile training ground for a graduate student and several undergraduate researchers in a wide variety of skills that range from computational methodology, experimental NMR and enzyme biochemistry. The undergraduates will be drawn from the pool of students who transfer from one of the seven community colleges that are part of the City University of New York (CUNY) into the City College of New York (CCNY) with a goal of encouraging them to pursue higher education in the STEM disciplines. It is notable that the graduate student who is expected on this project was formerly a transfer student who has chosen to pursue his PhD degree in Biochemistry. The transfer of a phosphate group from adenosine triphosphate (ATP) to specific serine, threonine or tyrosine residues of substrate proteins represents a central mechanism of signal transduction within cells and plays a critical role in almost all physiological processes. This transfer is enabled by biocatalysts known as protein kinases and involves the formation of a pre-chemistry complex between the kinase, the substrate and ATP. Recent studies suggest that the nature of the structural dynamics of the catalytic elements of the kinase and the phospho-acceptor region of the substrate in the context of this pre-chemistry complex determines the efficiency of the phospho-transfer reaction and contributes to its variability between specific kinase/substrate pairs. These variations in phosphorylation efficiency between kinase/substrate pairs is necessary to maintain the finely tuned network of numerous phosphorylation based signaling events occurring simultaneously in the cell. An understanding of these differences in efficiency requires the characterization in atomic detail of the conformational state/states that comprise the pre-chemistry complex. This project, using a combination of cutting-edge computational approaches, solution-state nuclear magnetic resonance (NMR) methods and biochemical assays, aims to characterize the conformational states that define the pre-chemistry complex towards the goal of addressing the following question: what determines the efficiency of phosphorylation within a kinase-substrate-ATP pre-chemistry complex? The model system chosen for this project involves the mitogen-activated protein kinase ERK2 and peptides derived from its natural substrate, the transcription factor Elk-1, that are phosphorylated with differing efficiencies.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

激酶是改变细胞中特定化学反应速率的生物分子。它们及这些化学反应的产物代表了生命所有领域的细胞内信号传导网络中的中心枢纽。除了细胞的定位和时间表达外，特定激酶对特定分子产物的效率对于将输入信号靶向不同的生理输出是关键。该项目的目的是通过在时空和时间上有足够的分辨率来理解该法规的性质，以提供驱动和最终设计信号网络的手段，这些信号网络将特定的输入信号驱动到定义的蜂窝响应。该项目还将为研究生和几位本科研究人员提供肥沃的培训场，包括计算方法论，实验性NMR和酶生物化学的各种技能。本科生将从纽约市城市大学（CUNY）的七个社区学院之一转移到纽约市学院（CCNY）的学生中，以鼓励他们在STEM学科中攻读高等教育。值得注意的是，在这个项目上期望的研究生以前是转学专业的学生，​​他选择攻读生物化学博士学位。 从三磷酸腺苷（ATP）转移到底物蛋白的特定丝氨酸，苏氨酸或酪氨酸残基，代表了细胞内信号转导的核心机制，并且在几乎所有生理过程中都起着至关重要的作用。这种转移由称为蛋白激酶的生物催化剂启用，涉及激酶，底物和ATP之间的化学络合物形成。最近的研究表明，在该预化学络合物的背景下，激酶催化元件的结构动力学和底物的磷酸化元素区域的性质决定了磷酸转移反应的效率，并有助于其在特定激酶/底物对之间的变异性。对于保持细胞中同时发生的许多基于磷酸化的信号事件的细胞调节网络，需要进行激酶/底物对之间磷酸化效率的这些变化。对效率上的这些差异的理解需要在构象状态/状态的原子细节中进行表征，该状态构成了化学综合体。该项目结合了尖端计算方法，解决方案状态核磁共振（NMR）方法和生化测定，旨在表征定义前化学综合体的构象状态，以解决以下问题的目的：解决以下问题：是什么决定了磷酸化的磷酸化效率在动力学酶-substrate-attpe-atpre-Atp-prepstrep-aTp-chemprate-atempp-chemprate-chemprate-atemistry中？为该项目选择的模型系统涉及有丝分裂原激活的蛋白激酶ERK2和肽源自其自然底物（转录因子ELK-1），这些转录因子ELK-1，这些因素ELK-1，其效率不同。这奖反映了NSF的法定任务，并通过使用基础的智力效果和宽阔的范围进行评估，并被认为值得评估。

项目成果

Nature of the Pre-Chemistry Ensemble in Mitogen-Activated Protein Kinases

• DOI: 10.1016/j.jmb.2018.12.007
• 发表时间: 2019-01-18
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Ghose, Ranajeet

Long-range dynamic correlations regulate the catalytic activity of the bacterial tyrosine kinase Wzc

• DOI: 10.1126/sciadv.abd3718
• 发表时间: 2020-12-01
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Hajredini, Fatlum;Piserchio, Andrea;Ghose, Ranajeet
• 通讯作者: Ghose, Ranajeet