Allosteric modulation of EAG1 gating

EAG1 门控的变构调节

• 批准号: 8421171
• 负责人: Gail A Robertson
• 金额: $ 31.04万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8421171
• 关键词: Address; Adopted; Alanine; Amino Acids; Behavior; Binding; Biochemical; Biological Assay; Brain; Calorimetry; Cancerous; Cells; Chemical Modifier; Chemicals; Complex; Coupling; Cyclic AMP-Dependent Protein Kinases; Cyclic Nucleotides; Cysteine; Data; Enzymes; Exhibits; Family; Family member; Fluorescent Probes; Fluorometry; In Vitro; Investigation; Ion Channel; Knowledge; Laboratories; Lead; Ligands; Link; Malignant Neoplasms; Mediating; Methodology; Modeling; Molecular Conformation; Monitor; Movement; Mutagenesis; N-terminal; Neuraxis; Outcome; Pharmaceutical Preparations; Physiological; Proteins; Reagent; Regulation; Reporting; Roentgen Rays; Role; Screening procedure; Side; Structure; Surface Plasmon Resonance; System; Testing; Therapeutic; Voltage-Gated Potassium Channel; Work; Xenopus oocyte; chemotherapy; cyclic-nucleotide gated ion channels; deletion analysis; drug development; in vitro testing; loss of function; molecular marker; mutant; novel; research study; sensor; therapeutic development; tool; voltage; voltage clamp

DESCRIPTION (provided by applicant): Eag-related channels (EAG, hERG, ELK) all share a highly conserved region bearing homology with cyclic- nucleotide binding domains of a variety of other proteins including cyclic nucleotide-gated channels. However, most evidence indicates this domain has evolved to serve a different, but unknown, function in Eag-related channels. This application introduces a new X-ray crystal structure for the EAG1 cyclic nucleotide homology domain (CNBhD) and proposes experiments directed by structural considerations to probe its function as an allosteric modulator of channel gating. Molecules interacting with this domain and modifying channel behavior will be identified through a layered screening approach. A range of biochemical and functional approaches reflecting complementary strengths of the participating laboratories will be employed. This project is expected to address a long-standing question in the field regarding the structure and function of CNBhD's, and produce reagents that will ultimately help define the functional role of EAG1 channels in the brain and facilitate chemotherapeutic drug development. PUBLIC HEALTH RELEVANCE: EAG1 channels are a major component of the electrical machinery of the brain and are also highly expressed in cancerous cells outside the brain, making them a potential target for chemotherapy. This proposal seeks to understand the inner workings of channel behavior, which are currently poorly understood. This new knowledge, combined with the proposed identification of new chemicals that can interfere or enhance channel function, may ultimately help define EAG1 function in the brain and facilitate the development of therapeutic drugs.

描述（由申请人提供）：Eag相关通道（EAG、hERG、ELK）都共享一个高度保守的区域，该区域与多种其他蛋白质（包括环核苷酸门控通道）的环核苷酸结合域具有同源性。然而，大多数证据表明，该结构域已进化为在 Eag 相关通道中发挥不同但未知的功能。该申请引入了 EAG1 环核苷酸同源结构域 (CNBhD) 的新 X 射线晶体结构，并提出了以结构考虑为指导的实验，以探讨其作为通道门控变构调节剂的功能。与该域相互作用并改变通道行为的分子将通过分层筛选方法来识别。将采用一系列反映参与实验室互补优势的生化和功能方法。该项目预计将解决该领域长期存在的有关 CNBhD 结构和功能的问题，并生产最终有助于确定 EAG1 通道在大脑中的功能作用并促进化疗药物开发的试剂。 公共健康相关性：EAG1 通道是大脑电机的主要组成部分，并且在脑外癌细胞中也高度表达，使其成为化疗的潜在靶点。该提案旨在了解目前人们对渠道行为的内部运作知之甚少。这些新知识，再加上拟议鉴定的能够干扰或增强通道功能的新化学物质，最终可能有助于确定大脑中 EAG1 的功能，并促进治疗药物的开发。