ERK2: Structure, Function and Inhibition

ERK2：结构、功能和抑制

基本信息

批准号：
8130662
负责人：
Kevin N Dalby
金额：
$ 32.33万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-04-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8130662
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 MAPK4 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 Phosphotransferases Principal Investigator Process Protein Isoforms Protein Kinase Proteins Recruitment Activity Regulation Research Roentgen Rays S Phase 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 enzyme substrate complex extracellular human disease inhibitor/antagonist insight member migration mouse model peptidomimetics pre-clinical prevent programs prospective protein protein interaction response small molecule stress-activated protein kinase 1 therapeutic target translational study

项目摘要

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（由申请人提供）：BRAF致癌突变发生在大约8％的人类癌症中，并导致Ser/Thr特异性蛋白激酶ERK1和ERK2的持续活性，它们是细胞进入人类细胞周期S相的关键介体。为了开始理解ERK1/2的底物识别和催化的原理，将使用一系列复杂的NMR技术在AIM 1中确定ERK27ETS1（酶7substrate）复合物的结构和动力学。转录因子ELK-1（ERK1/2的另一个底物）的半合成双基层类似物将构建以模拟磷酸化转移，并以类似的方式进行分析。这些研究将通过X射线晶体结构方法来称赞。在AIM 2中，通过大大增加共识结合序列中的侧链多样性，可以获得有效和高度选择性的靶向分子，这些分子将通过ERK1/2及其蛋白质配体之间的相互作用。这些靶向分子的作用机理和特异性将使用酶动力学和结构方法确定。在AIM 3中，靶向分子将使用蛋白质转导域将其输送到细胞中，并根据其抑制生长因子刺激的ERK1/2活性和细胞生长的能力确定其功效。值得注意的是，该跨学科的研究计划将首次提供ERK1/2的有效抑制剂，并允许ERK1/2底物恢复地点被证实为人类癌症的潜在药物靶标。本文开发的方法将迅速适用于其他蛋白激酶，从而潜在地包含许多人类疾病，并为未来的临床小鼠模型中的未来翻译研究提供基础。公共卫生相关性：该应用的重点是一种称为ERK2的酶，其活动在大量的人类癌症中被上调。拟议的研究将提供有关该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鉴定可阻止ERK2结合其他细胞蛋白的可渗透分子。这将在人类疾病模型中提供ERK2作为药物靶标的验证，并为验证同一酶家族的其他成员提供实验基础。提出的研究将具有很高的意义，并将有益于人类健康。

项目成果

期刊论文数量（40）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mechanistic Studies on Covalent Assemblies of Metal-Mediated Hemi-Aminal Ethers.

DOI：
10.1039/c4sc02495h
发表时间：
2015-01-01
期刊：
Chemical science
影响因子：
8.4
作者：
Jo HH;Edupuganti R;You L;Dalby KN;Anslyn EV
通讯作者：
Anslyn EV

Examining docking interactions on ERK2 with modular peptide substrates.

DOI：
10.1021/bi201103b
发表时间：
2011-11-08
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Lee, Sunbae;Warthaka, Mangalika;Yan, Chunli;Kaoud, Tamer S.;Ren, Pengyu;Dalby, Kevin N.
通讯作者：
Dalby, Kevin N.

Computational insights for the discovery of non-ATP competitive inhibitors of MAP kinases.

发现 MAP 激酶的非 ATP 竞争性抑制剂的计算见解。

DOI：
10.2174/138161212799436368
发表时间：
2012
期刊：
Current pharmaceutical design
影响因子：
3.1
作者：
Schnieders,MichaelJ;Kaoud,TamerS;Yan,Chunli;Dalby,KevinN;Ren,Pengyu
通讯作者：
Ren,Pengyu

In-situ generation of differential sensors that fingerprint kinases and the cellular response to their expression.

DOI：
10.1021/ja407397z
发表时间：
2013-10-02
期刊：
Journal of the American Chemical Society
影响因子：
15
作者：
Zamora-Olivares D;Kaoud TS;Dalby KN;Anslyn EV
通讯作者：
Anslyn EV

Virtual screening using molecular simulations.

DOI：
10.1002/prot.23018
发表时间：
2011-06
期刊：
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
影响因子：
2.9
作者：
Yang, Tianyi;Wu, Johnny C.;Yan, Chunli;Wang, Yuanfeng;Luo, Ray;Gonzales, Michael B.;Dalby, Kevin N.;Ren, Pengyu
通讯作者：
Ren, Pengyu