ERK2: Structure, Function and Inhibition

ERK2：结构、功能和抑制

• 批准号: 7658699
• 负责人: Kevin N Dalby
• 金额: $ 32.92万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7658699
• 关键词: Apoptosis; BRAF gene; Binding; Biological; Cancer cell line; Carboxylic Acids; Catalysis; Cell Cycle; Cell fusion; Cells; Clinical; Communication; Complement; Complex; Consensus; Cyclin D1; Data; Degenerative Disorder; Development; Discipline; Disease; Doctor of Philosophy; Drug Delivery Systems; Enzyme Kinetics; Enzymes; Event; FOS gene; Family; Funding; Future; Genes; Goals; Growth; Growth Factor; Health; Human; Immediate-Early Genes; In Vitro; Individual; Kinetics; Lead; Libraries; Life; Ligands; MAP Kinase Gene; MAPK1 gene; MAPK10 gene; MAPK14 gene; MAPK3 gene; MAPK7 gene; MAPK8 gene; MAPK9 gene; Malignant Neoplasms; Mediator of activation protein; Methodology; Methods; Mitogen-Activated Protein Kinases; Mitogens; Modeling; Mus; Mutation; Nature; Nuclear; Oncogenic; Peptides; Pharmaceutical Preparations; Phase; Phosphotransferases; Principal Investigator; Process; Protein Isoforms; Protein Kinase; Proteins; Recruitment Activity; Regulation; Research; Roentgen Rays; Scanning; Side; Signal Transduction; Site; Specificity; Structure; Substrate Interaction; Substrate Specificity; System; Techniques; Therapeutic; Time; Tissues; Toxic effect; Validation; X-Ray Crystallography; analog; base; cancer therapy; cell growth; elk-1 protein; extracellular; human disease; inhibitor/antagonist; insight; member; migration; mouse model; peptidomimetics; pre-clinical; prevent; programs; prospective; protein protein interaction; public health relevance; response; small molecule; stress-activated protein kinase 1; therapeutic target; translational study

DESCRIPTION (provided by applicant): BRAF oncogenic mutations occur in approximately 8% of human cancers and lead to sustained activity of the Ser/Thr-specific protein kinases ERK1 and ERK2, which are critical mediators of cell entry into the S- phase of the human cell cycle. To begin to understand the principles governing substrate recognition and catalysis by ERK1/2, the structure and dynamics of the ERK27Ets1 (enzyme7substrate) complex will be determined in Aim 1, using an array of sophisticated NMR techniques. A semi-synthetic bisubstrate analog of transcription factor Elk-1, another substrate of ERK1/2, will be constructed to mimic phosphoryl transfer and analyzed in a similar manner. These studies will be complimented by X-ray crystal structure approaches. In Aim 2, potent and highly selective Targeting Molecules that block interactions between ERK1/2 and its protein ligands will be acquired by greatly increasing the side chain diversity within consensus binding sequences. The mechanism of action and specificity of these targeting molecules will be determined using enzyme kinetics and structural approaches. In Aim 3, the Targeting Molecules will be delivered into cells using protein transduction domains, and their efficacy determined with respect to their ability to inhibit Growth Factor-stimulated ERK1/2 activity and cell growth. Significantly, this multi-disciplined research program will provide potent and specific inhibitors of ERK1/2 for the first time and will allow ERK1/2 substrate-recruiting sites to be validated as potential drug targets for human cancers. The methodology developed herein will be rapidly applicable to other protein kinases, thereby potentially embracing many human diseases, and will provide the basis for future translational studies in pre-clinical mouse models. PUBLIC HEALTH RELEVANCE: The focus of this application is an enzyme called ERK2 whose activity is upregulated in a large number of human cancers. The proposed studies will provide detailed structural information on how this enzyme interacts with other cellular proteins. A new technique will be developed to rapidly identify cell-permeable molecules that can prevent ERK2 from binding other cellular proteins. This will provide validation of ERK2 as a drug target, in models of human disease, and provide the experimental basis for validating other members of the same enzyme family. The studies proposed will be of high significance and will benefit human health.

描述（由申请方提供）：BRAF致癌突变发生在约8%的人类癌症中，并导致Ser/Thr特异性蛋白激酶ERK 1和ERK 2的持续活性，这是细胞进入人类细胞周期S期的关键介导物。为了开始理解ERK 1/2的底物识别和催化原理，我们将在目标1中使用一系列复杂的NMR技术来确定ERK 27 Ets 1（酶7底物）复合物的结构和动力学。将构建转录因子Elk-1（ERK 1/2的另一种底物）的半合成双底物类似物以模拟磷酰基转移，并以类似方式进行分析。这些研究将通过X射线晶体结构方法得到补充。在目标2中，将通过大大增加共有结合序列内的侧链多样性来获得阻断ERK 1/2与其蛋白配体之间相互作用的有效且高选择性的靶向分子。这些靶向分子的作用机制和特异性将使用酶动力学和结构方法来确定。在目的3中，靶向分子将使用蛋白转导结构域递送到细胞中，并且它们的功效根据它们抑制生长因子刺激的ERK 1/2活性和细胞生长的能力来确定。值得注意的是，这项多学科的研究计划将首次提供有效和特异性的ERK 1/2抑制剂，并将允许ERK 1/2底物招募位点被验证为人类癌症的潜在药物靶点。本文开发的方法将迅速适用于其他蛋白激酶，从而可能涵盖许多人类疾病，并将为未来临床前小鼠模型的翻译研究提供基础。公共卫生相关性：该应用的重点是一种名为ERK 2的酶，其活性在大量人类癌症中上调。拟议的研究将提供有关这种酶如何与其他细胞蛋白相互作用的详细结构信息。将开发一种新技术来快速识别可阻止ERK 2结合其他细胞蛋白的细胞渗透性分子。这将在人类疾病模型中验证ERK 2作为药物靶点，并为验证同一酶家族的其他成员提供实验基础。所提议的研究将具有重要意义，并将有益于人类健康。