Physiologically relevant energetic interactions within the Na/K Pump

Na/K 泵内生理相关的能量相互作用

• 批准号: 8574225
• 负责人: Pablo Artigas
• 金额: $ 45.26万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8574225
• 关键词: ATP phosphohydrolase; Active Biological Transport; Address; Affinity; Amino Acids; Animals; Arrhythmia; Binding; Binding Sites; Biochemistry; Brain; Cardiac Glycosides; Cardiovascular Diseases; Carrier Proteins; Cell membrane; Cells; Characteristics; Chemicals; Complex; Computer Simulation; Congestive Heart Failure; Cysteine; Data; Digitalis preparation; Disease; Disulfides; Drug usage; Electrodes; Electrophysiology (science); Engineering; Etiology; Familial Hemiplegic Migraine; Familial disease; Functional disorder; Gene Mutation; Genes; Goals; Heart failure; Hypertension; Investigation; Ions; Laboratories; Lead; Life; Ligands; Link; Maintenance; Manuscripts; Mediating; Membrane Potentials; Membrane Proteins; Migraine; Mission; Modification; Molecular; Molecular Biology; Molecular Conformation; Muscle; Mutagenesis; Mutate; Mutation; Myocardium; Na(+)-K(+)-Exchanging ATPase; Nerve; Oocytes; Parkinsonian Disorders; Pharmaceutical Preparations; Polycystic Kidney Diseases; Positioning Attribute; Protein Isoforms; Pump; Recruitment Activity; Research; Resolution; Rest; Role; Structural Models; Structure; Students; Surface; Testing; Tissues; Work; Xenopus oocyte; base; clinically relevant; computational chemistry; conformer; crosslink; extracellular; graduate student; inhibitor/antagonist; insight; interest; meetings; molecular mechanics; mutant; nervous system disorder; novel; palytoxin; patch clamp; public health relevance; receptor; single molecule; undergraduate student; voltage clamp

DESCRIPTION (provided by applicant): All animal cells require maintenance of transmembrane gradients for Na+ and K+ ions by the Na/K pump, a plasma membrane P-type ATPase. Pathophysiologically, the Na/K pump is the target specifically inhibited by cardiotonic steroids, drugs traditionally used for the treatment of heart failure. Also, mutations in the genes encoding for two Na/K pump isoforms have been linked to migraine and Parkinsonism. The long-term goal of our laboratory is to understand the relationship between the Na/K pump structure, its function, and its multiple roles in cardiovascular and neurological diseases. All P-type ATPases alternate between two major conformers E1 and E2. The molecular forces that stabilize intermediate states in the cycle must arise as a consequence of a set of state-specific residue-residue and residue-ligand interactions. The main research goal of this AREA project is to identify and characterize energetically and mechanistically relevant interactions between pairs of residues and between residues and ions within the Na/K pump. To address this problem we use mutagenesis, heterologous expression of Na/K pumps in Xenopus oocytes, voltage clamp (two electrode voltage clamp, cut-open oocyte and patch clamp), chemical modification of engineered cysteine residues and computational chemistry to address two independent and interrelated aims, which we propose on the basis of extensive preliminary work: 1) To identify and characterize critical conformation-specific interactions that stabilize Na/K pump conformations, 2) to elucidate the mechanisms of ion-induced conformational changes. Considering the overwhelming evidence that the rate of E1 - E2 conversion underlies the molecular mechanism of this critical transporter, the work proposed here will be crucial to understanding and resolving the etiology of clinically relevant problems. Specifically, dysfunction of the Na,K-ATPase has been attributed to hypertension, congestive heart failure, familial hemiplegic migraine, and polycystic kidney disease.

描述（由申请人提供）：所有动物细胞都需要通过Na/K泵（一种质膜p型atp酶）维持Na+和K+离子的跨膜梯度。病理生理上，Na/K泵是被强心剂特异性抑制的靶点，而强心剂是传统上用于治疗心力衰竭的药物。还有，基因突变

项目成果

Displacement of the Na + /K + pump’s transmembrane domains demonstrates conserved conformational changes in P-type 2 ATPases

Na /K 泵跨膜结构域的置换证明了 P 型 2 ATP 酶中保守的构象变化

• DOI: 10.1073/pnas.2019317118
• 发表时间: 2021
• 期刊: Proceedings of the National Academy of Sciences
• 作者: Young, Victoria C.;Artigas, Pablo
• 通讯作者: Artigas, Pablo