Investigating KA1 domain-mediated regulation of MARK/PAR1 kinases

研究 KA1 结构域介导的 MARK/PAR1 激酶调节

• 批准号: 9062868
• 负责人: Ryan P. Emptage
• 金额: $ 5.61万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9062868
• 关键词: Address; Adverse effects; Affinity; Allosteric Regulation; Alzheimer' s Disease; Amino Acids; Autistic Disorder; Back; Binding; Binding Proteins; C-terminal; Cell physiology; Cells; Development; Disease; Distant; Drug Design; Exposure to; F2R gene; Goals; Health; Heart Diseases; Homologous Gene; Kinetics; Lead; Learning; Length; Ligands; Light; Link; Lipids; MAPT gene; Malignant Neoplasms; Mediating; Membrane; Methods; Microtubules; Molecular; Mutation; Oranges; Organism; Pathology; Phospholipids; Phosphotransferases; Play; Protein Kinase; Proteins; Regulation; Role; Scaffolding Protein; Signal Transduction; Site; Stimulus; Substrate Specificity; Tail; Testing; UBA Domain; Work; base; biophysical techniques; design; enzyme activity; in vitro activity; in vivo; inhibitor/antagonist; inorganic phosphate; insight; kinase inhibitor

 DESCRIPTION (provided by applicant): MARK/PAR1 kinases occupy critical signaling nodes in the development of higher organisms, and have been linked to a plethora of disease states including Autism, Alzheimer's, cancer, and heart disease. The goal of this proposal is to elucidate the structural and functional role of the kinase-associated 1 (KA1) domain, which resides at the C-terminus of MARK/PAR1 and related kinases. This module has recently been shown to interact with both phospholipid and protein ligands, but has also been proposed to provide an autoinhibitory function for the attached kinase domain. Preliminary studies have established that a separately purified KA1 module does bind the MARK1 kinase domain and can inhibit activity in vitro. We hypothesize that kinase autoinhibition is relieved upon binding o the KA1 domain to lipid and/or protein ligands and plan to rigorously assess these features of MARK/PAR1 kinases. The aims of this project are to (i) fully characterize the interaction between the MARK1 kinase and KA1 domains through a battery of structural and biophysical methods and, and (ii) establish a functional role for the KA1 domain in MARK1 activity by in vitro kinetic and in vivo cell-based methods, including determination of whether exposure to lipid or protein ligands of this domain disrupt autoinhibition and activate the kinase. These studies will shed light onto how varied inputs from lipid or protein ligands can lead to kinase regulation, and will provide the groundwork for design of specific allosteric inhibitors of MARK/PAR1 kinases.

 描述（由适用提供）：Mark/Par1激酶在更高生物体的发展中占据关键的信号传导节点，并且与包括自闭症，阿尔茨海默氏症，癌症和心脏病在内的众多疾病状态有关。该提案的目的是阐明激酶相关1（KA1）结构域的结构和功能作用，该结构域在Mark/PAR1和相关激酶的C端住宅。该模块最近已显示与磷脂和蛋白质配体相互作用，但也已提出为附着的激酶结构域提供自身抑制功能。初步研究已经确定，单独纯化的KA1模块确实结合了Mark1激酶结构域并可以在体外抑制活性。我们假设激酶自抑制在结合Ka1结构域与脂质和/或蛋白质配体的结合后可以放弃，并计划严格评估Mark/Par1激酶的这些特征。 The aims of this project are to (i) fully characterize the interaction between the MARK1 kinase and KA1 domains through a battery of structural and biophysical methods and, and (ii) establish a functional role for the KA1 domain in MARK1 activity by in vitro kinetic and in vivo cell-based methods, including determination of whether exposure to lipid or protein ligands of this domain disrupt autoinhibition and activate the激酶。研究将阐明脂质或蛋白质配体的不同输入会导致激酶调节，并为设计特定的Mark/Par1激酶的特定变构抑制剂设计提供基础。