The circadian clock protein BMAL and post-translational regulation of ENaC in the kidney

肾脏中生物钟蛋白 BMAL 和 ENaC 的翻译后调节

• 批准号: 10662317
• 负责人: Abdel Ayube Alli
• 金额: $ 33.55万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662317
• 关键词: ARNTL gene; Adaptor Signaling Protein; Address; Aldosterone; Antihypertensive Agents; Apical; Attenuated; Binding; Biological Assay; Biotinylation; Blood Pressure; Cardiovascular Diseases; Caspase; Cathepsins B; Cause of Death; Cell membrane; Cells; Clock protein; Data; Distal; Duct (organ) structure; Ductal Epithelial Cell; Electrolytes; Equilibrium; Essential Hypertension; Etiology; Excretory function; Exhibits; Female; Fluorescence Resonance Energy Transfer; Genes; Goals; Grant; Hour; Hypertension; In Situ; Infusion procedures; Investigation; Ion Channel; Kidney; Knockout Mice; Knowledge; MARCKS gene; Mass Spectrum Analysis; Measures; Mediating; Membrane; Metabolic; Molecular; Mus; Nephrons; Patch-Clamp Techniques; Pathogenesis; Peptide Hydrolases; Phase; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipids; Play; Post-Translational Protein Processing; Post-Translational Regulation; Potassium; Probability; Proteins; Proteolysis; Proteome; Proteomics; Regulation; Role; Sex Differences; Sodium; Telemetry; Testing; Therapeutic; Time; United States; Up-Regulation; Western Blotting; Wild Type Mouse; absorption; alpha 1-Antitrypsin; apical membrane; blood pressure control; blood pressure reduction; blood pressure regulation; cardiovascular risk factor; circadian; circadian pacemaker; circadian regulation; comparison control; density; epithelial Na+ channel; experimental study; male; new therapeutic target; novel; overexpression; protein expression; renal epithelium; salt balance; sodium channel proteins; transcription factor; urinary; ARNTL gene; Adaptor Signaling Protein; Address; Aldosterone; Antihypertensive Agents; Apical; Attenuated; Binding; Biological Assay; Biotinylation; Blood Pressure; Cardiovascular Diseases; Caspase; Cathepsins B; Cause of Death; Cell membrane; Cells; Clock protein; Data; Distal; Duct (organ) structure; Ductal Epithelial Cell; Electrolytes; Equilibrium; Essential Hypertension; Etiology; Excretory function; Exhibits; Female; Fluorescence Resonance Energy Transfer; Genes; Goals; Grant; Hour; Hypertension; In Situ; Infusion procedures; Investigation; Ion Channel; Kidney; Knockout Mice; Knowledge; MARCKS gene; Mass Spectrum Analysis; Measures; Mediating; Membrane; Metabolic; Molecular; Mus; Nephrons; Patch-Clamp Techniques; Pathogenesis; Peptide Hydrolases; Phase; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipids; Play; Post-Translational Protein Processing; Post-Translational Regulation; Potassium; Probability; Proteins; Proteolysis; Proteome; Proteomics; Regulation; Role; Sex Differences; Sodium; Telemetry; Testing; Therapeutic; Time; United States; Up-Regulation; Western Blotting; Wild Type Mouse; absorption; alpha 1-Antitrypsin; apical membrane; blood pressure control; blood pressure reduction; blood pressure regulation; cardiovascular risk factor; circadian; circadian pacemaker; circadian regulation; comparison control; density; epithelial Na+ channel; experimental study; male; new therapeutic target; novel; overexpression; protein expression; renal epithelium; salt balance; sodium channel proteins; transcription factor; urinary

ABSTRACT The epithelial sodium channel (ENaC) expressed in the distal tubule and collecting duct is responsible for the final regulation of sodium reabsorption by the kidneys. The myristoylated alanine-rich C kinase substrate (MARCKS) plays an important role as an adaptor protein between the anionic phospholipid PIP2 and ENaC. Both ENaC and MARCKS are positively regulated by the protease cathepsin B. First, our preliminary data demonstrate renal ENaC activity and MARCKS protein expression are positively regulated by the circadian protein BMAL1. Second, our preliminary data show alpha-1 antitrypsin is increased in the BMAL1 knockout mouse kidney compared to the kidney of wild-type mice sacrificed at the same time. Third, our preliminary data show alpha-1 antitrypsin is expressed in the kidney and it strongly inhibits cathepsin B activity and contributes to blood pressure regulation. In this project we will test our hypothesis that the association between renal ENaC and MARCKS, and their function at the apical plasma membrane negatively correlates with alpha-1 antitrypsin expression in a circadian dependent manner. We will perform experiments to investigate proteolysis and apical membrane expression of ENaC and MARCKS, ENaC activity, sodium handling, and blood pressure using male and female BMAL1 knockout mice, alpha-1 antitrypsin knockout mice, alpha-1 antitrypsin overexpressing mice, cathepsin B knockout mice, and wild-type control mice. The successful completion of our proposed studies for this project will reveal new mechanisms underlying the role of BMAL1 in the regulation of renal ENaC and MARCKS and blood pressure control. Our long term goal is to provide a better understanding for the pathogenesis of essential hypertension that can potentially lead to novel drug targets and therapeutics.

抽象的 在远端小管中表达并收集管道的上皮钠通道（ENAC） 负责最终调节肾脏钠的重吸收。这 肉豆蔻酰化富含丙氨酸的C激酶底物（MARCKS）作为一个重要作用 阴离子磷脂PIP2和ENAC之间的衔接蛋白。 enac和 马克克人受蛋白酶组织蛋白酶B的积极调节。首先，我们的初步 数据表明肾脏ENAC活性和MARCKS蛋白表达是积极的 由昼夜节律BMAL1调节。第二，我们的初步数据显示Alpha-1 与肾脏相比 野生型小鼠同时牺牲了。第三，我们的初步数据显示Alpha-1 抗胰蛋白酶在肾脏中表达，并强烈抑制组织蛋白酶B活性和 有助于血压调节。在这个项目中，我们将测试我们的假设 肾脏ENAC和MARCKS之间的关联及其在顶端的功能 质膜与A中的α-1抗胰蛋白酶表达负相关 昼夜节律的方式。我们将进行实验以研究蛋白水解 ENAC和MARCK的顶膜表达，ENAC活性，钠 使用雄性和雌性BMAL1基因敲除小鼠的处理和血压，Alpha-1 抗胰蛋白酶基因敲除小鼠，α-1抗胰蛋白酶过表达小鼠，组织蛋白酶B 敲除小鼠和野生型对照小鼠。我们提议的成功完成 该项目的研究将揭示BMAL1在 调节肾脏ENAC和MARCKS以及血压控制。我们的长期目标 是为基本高血压的发病机理提供更好的理解 可能会导致新颖的药物靶标和治疗剂。