Potassium Channels as Macromolecular Complexes

作为大分子复合物的钾通道

• 批准号: 8741656
• 负责人: William A Coetzee
• 金额: $ 28.96万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-27 至 2015-09-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8741656
• 关键词: Affect; Aldehyde-Lyases; Biochemistry; Biological Assay; Blood flow; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Co-Immunoprecipitations; Complement; Complex; Coronary; Data; Depressed mood; Dilatation - action; Disease; Dominant-Negative Mutation; Electrophysiology (science); Endothelium; Energy Metabolism; Enzyme Interaction; Enzymes; Event; Exercise; Functional disorder; Genetic; Glycolysis; Health; Heart; Hypertrophy; Hypoxia; Impairment; Individual; Ion Channel; Ischemia; Knowledge; Link; Macromolecular Complexes; Membrane; Metabolic; Modality; Molecular; Molecular Biology; Muscle Cells; Myocardial; Myocardial Ischemia; Myocardium; Nucleotides; Pathway interactions; Physiological; Physiological Processes; Potassium; Potassium Channel; Proteins; Proteomics; Pyruvate Kinase; Regulation; Research; Role; Smooth Muscle; Specificity; Stimulus; Stress; Techniques; Testing; Tissues; Transgenic Mice; Triose-Phosphate Isomerase; Ventricular; biological adaptation to stress; cell type; glucose metabolism; insight; insulin secretion; interest; mouse model; mutant; novel; prevent; promoter; protective effect; protein complex; reactive hyperemia; research study; response; tool

DESCRIPTION (provided by applicant): ATP-sensitive potassium [KATP] channels in the heart muscle and coronary myocytes couple cellular metabolic status to membrane excitability, thereby contributing to the regulation of tissue responses to physiological and pathophysiological stimuli. In the heart muscle, opening of KATP channels participate in the stress response and protect against ischemic episodes. In the coronary vasculature, K(ATP/NDP) channels contribute to the regulation of basal flow as well as responses to metabolic impairment (hypoxic dilatation and ischemic reactive hyperemia). We found glycolytic enzymes to associate with KATP channel subunits, We hypothesize that glycolytic enzymes are integral components of the KATP channel macromolecular complex and that glycolytic enzymes regulate KATP channel activity under physiological and pathophysiological conditions, both in the cardiac myocyte as well as in the coronary smooth muscle and endothelium. In a first Specific Aim, we will investigate the hypothesis that enzymes of the glycolytic complex are associated with the KATP channel. Using co-immunoprecipitation assays we will investigate the specificity of interaction of glycolytic enzymes with individual KATP channel subunits (Kir6.1, Kir6.2, SUR1, SUR2A and SUR2B). Co-immunoprecipitation assays of native proteins will be performed to investigate interactions under physiologically relevant conditions. Protein interactions will also be investigated using advanced proteomic approaches (ICAT & ITRAQ), which has the potential to uncover additional novel KATP channel interacting proteins. In a second Aim, we will examine the hypothesis that physical interaction of glycolytic enzymes with KATP channel subunits is required and that channel modulation occurs because of altered nucleotide levels in the microenvironment of the channel complex. This will be accomplished using mutant KATP channel subunits (lacking interaction with glycolytic enzymes or altered nucleotide sensitivity). In a final Aim, we will investigate the interaction of glycolysis and KATP channels in the context of ischemic protection in cardiac myocytes and the coronary vasculature. To this end, we will utilize our genetic mouse models that express dominant-negative K(ATP) channel subunits specifically in the cardiac myocyte, smooth muscle or endothelium. Our findings may have important implications for understanding the role of KATP channels in the heart and coronary vasculature under physiological and non-pathophysiological conditions.

描述（由申请人提供）：心肌和冠状肌细胞中的 ATP 敏感钾 [KATP] 通道将细胞代谢状态与膜兴奋性耦合，从而有助于调节组织对生理和病理生理刺激的反应。在心肌中，KATP 通道的开放参与应激反应并防止缺血发作。在冠状脉管系统中，K(ATP/NDP) 通道有助于调节基础流量以及对代谢损伤（缺氧扩张和缺血反应性充血）的反应。我们发现糖酵解酶与 KATP 通道亚基相关，我们假设糖酵解酶是 KATP 通道大分子复合物的组成部分，并且糖酵解酶在生理和病理生理条件下调节心肌细胞和冠状动脉中的 KATP 通道活性平滑肌和内皮。在第一个特定目标中，我们将研究糖酵解复合物的酶与 KATP 通道相关的假设。使用免疫共沉淀测定，我们将研究糖酵解酶与单个 KATP 通道亚基（Kir6.1、Kir6.2、SUR1、SUR2A 和 SUR2B）相互作用的特异性。将进行天然蛋白质的免疫共沉淀测定，以研究生理相关条件下的相互作用。还将使用先进的蛋白质组学方法（ICAT 和 ITRAQ）研究蛋白质相互作用，该方法有可能发现其他新型 KATP 通道相互作用蛋白质。在第二个目标中，我们将检验以下假设：糖酵解酶与 KATP 通道亚基之间的物理相互作用是必需的，并且通道调节是由于通道复合体微环境中核苷酸水平的改变而发生的。这将通过使用突变的 KATP 通道亚基（缺乏与糖酵解酶的相互作用或改变的核苷酸敏感性）来实现。在最终目标中，我们将研究心肌细胞和冠状脉管系统缺血保护背景下糖酵解和 KATP 通道的相互作用。为此，我们将利用我们的遗传小鼠模型，该模型特异地在心肌细胞、平滑肌或内皮细胞中表达显性失活 K(ATP) 通道亚基。我们的研究结果可能对理解生理和非病理生理条件下 KATP 通道在心脏和冠状血管系统中的作用具有重要意义。

项目成果

Measuring and evaluating the role of ATP-sensitive K+ channels in cardiac muscle.

测量和评估 ATP 敏感 K 通道在心肌中的作用。

• DOI: 10.1016/j.yjmcc.2011.12.012
• 发表时间: 2012
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Kefaloyianni,Eirini;Bao,Li;Rindler,MichaelJ;Hong,Miyoun;Patel,Tejaskumar;Taskin,Eylem;Coetzee,WilliamA
• 通讯作者: Coetzee,WilliamA

Multiplicity of effectors of the cardioprotective agent, diazoxide.

• DOI: 10.1016/j.pharmthera.2013.06.007
• 发表时间: 2013-11
• 期刊: PHARMACOLOGY & THERAPEUTICS
• 影响因子: 13.5
• 作者: Coetzee, William A.