STRUCTURE, FUNCTION, AND REGULATION OF CASEIN KINASE-1

酪蛋白激酶-1 的结构、功能和调控

基本信息

批准号：
6386731
负责人：
Jeff Kuret
金额：
$ 20.05万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-08-01 至 2003-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6386731
关键词：
casein kinase chemical binding enzyme activity enzyme inhibitors enzyme structure enzyme substrate molecular cloning protein kinase protein purification site directed mutagenesis

项目摘要

DESCRIPTION (Adapted from applicant's abstract): The broad objective of this project is to deduce the structural basis of protein kinase ligand selectivity and regulation. The work focuses on the casein kinase-1 (CK1) family of enzymes, which are present in all eukaryotic cells and play an essential role in regulation of the cytoskeleton. In addition to their normal physiological significance, selected members of this enzyme family are leading candidates for mediating the pathological hyperphosphorylation of cytoskeletal proteins found in neurodegenerative diseases such as Alzheimer's disease. Although members of the CK1 family are numerous and comprise one of the largest branches of the protein kinase superfamily, preliminary data suggest commonality in their mechanism of protein substrate recognition, regulation by phosphorylation, and sensitivity to small-molecule inhibitors. The goal of this proposal is to test this hypothesis by a combination of biophysical and molecular biological methods. The specific aims are: (1) to establish the basis of CK1 substrate recognition and regulation. Guided by an existing CK1 crystal structure, specific models will be tested in vitro and in vivo using oligonucleotide-directed mutagenesis; (2) to isolate and characterize a novel CK1 regulatory kinase. An enzyme that phosphorylates a site homologous to a regulatory site in cyclin-dependent protein kinases will be purified, characterized biochemically, and cloned to establish its relationship to the cyclin-dependent kinase activating kinase (CAK); and (3) to discover the mechanism of action of novel small molecule inhibitors. The binding site occupied by these molecules will be identified and sought in other protein kinases of known 3-dimensional structure. Although these experiments focus on CK1, the results will have broad implications for the protein kinase family as whole. In addition, a complete understanding of protein kinase structure and inhibitor selectivity will speed the rational development of protein kinase-selective inhibitors with potentially useful therapeutic properties.

描述（改编自申请人的摘要）：该项目是推断蛋白激酶配体的结构基础选择性和调节。这项工作重点是酪蛋白激酶-1（CK1）酶家族，所有真核细胞中都存在并发挥作用在调节细胞骨架中的重要作用。除了他们正常的生理意义，该酶家族的选定成员是介导病理性磷酸化的领先候选者在神经退行性疾病中发现的细胞骨架蛋白，例如阿尔茨海默氏病。尽管CK1家族的成员很多，并且包括蛋白激酶超家族的最大分支之一初步数据表明其蛋白质底物的机理识别，磷酸化调节以及对小分子抑制剂。该提议的目的是测试通过生物物理和分子生物学方法的结合进行假设。具体目的是：（1）建立CK1底物的基础认可和法规。在现有的CK1晶体结构的指导下，特定模型将在体外和体内测试寡核苷酸指导的诱变；（2）隔离和表征新型CK1调节激酶。磷酸化位点的酶与细胞周期蛋白依赖性蛋白激酶中的调节位点同源纯化，生化特征并克隆以建立其与细胞周期蛋白依赖性激酶激活激酶（CAK）的关系；（3）发现新型小分子抑制剂的作用机理。这这些分子占据的结合位点将被鉴定并在已知三维结构的其他蛋白激酶。虽然这些实验以CK1为重点，结果将对蛋白激酶家族作为整体。此外，对蛋白激酶结构和抑制剂选择性将加快理性具有潜在有用的蛋白激酶选择性抑制剂的开发治疗特性。