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: 10440278
• 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/10440278
• 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; In Situ; Infusion procedures; Investigation; Ion Channel; Kidney; Knockout Mice; Knowledge; Lead; MARCKS gene; Mass Spectrum Analysis; Measures; Mediating; Membrane; Metabolic; Molecular; Mus; Nephrons; Patch-Clamp Techniques; Pathogenesis; Peptide Hydrolases; Pharmaceutical Preparations; 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; Time; United States; Up-Regulation; Western Blotting; Wild Type Mouse; alpha 1-Antitrypsin; apical membrane; blood pressure control; blood pressure reduction; blood pressure regulation; cardiovascular risk factor; circadian; circadian pacemaker; circadian regulation; density; epithelial Na+ channel; experimental study; hypertensive; male; new therapeutic target; novel; overexpression; protein expression; renal epithelium; salt balance; sodium channel proteins; targeted treatment; 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.

摘要 上皮性钠通道(ENaC)在远端肾小管和集合管的表达 负责最终调节肾脏对钠的重吸收。这个 肉豆蔻酰化富含丙氨酸的C激酶底物(MARCKS)作为一种 阴离子磷脂PIP2和ENaC之间的接头蛋白。ENaC和 Marcks受到组织蛋白酶B的正向调节。首先，我们的初步研究 数据显示肾脏ENaC活性和Marcks蛋白表达为正 受昼夜节律蛋白BMAL1调控。其次，我们的初步数据显示阿尔法-1 与BMAL1基因敲除小鼠肾脏相比，BMAL1基因敲除小鼠肾脏中抗胰蛋白酶活性增加 同时处死野生型小鼠。第三，我们的初步数据显示α-1 抗胰蛋白酶在肾脏中表达，它强烈抑制组织蛋白酶B的活性，并 有助于调节血压。在这个项目中，我们将测试我们的假设 肾脏ENaC和Marcks的相关性及其在心尖部的作用 细胞质膜与α-1抗胰蛋白酶表达呈负相关 昼夜节律依赖的方式。我们将进行实验来研究蛋白质的分解 ENaC和Marcks的表达、ENaC活性、钠 使用雄性和雌性BMAL1基因敲除小鼠α-1处理和血压 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 基因敲除小鼠和野生型对照小鼠。圆满完成我们提议的 该项目的研究将揭示BMAL1在 肾脏ENaC和Marcks的调节与血压控制。我们的长期目标 是为了更好地了解原发性高血压的发病机制 可能会导致新的药物靶点和治疗方法。