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

基本信息

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
项目类别：
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

项目摘要

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。