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.

抽象的上皮钠通道 (ENaC) 在远端小管和集合管中表达负责肾脏钠重吸收的最终调节。这肉豆蔻酰化富含丙氨酸的 C 激酶底物 (MARCKS) 作为阴离子磷脂 PIP2 和 ENaC 之间的衔接蛋白。 ENaC 和 MARCKS 受到蛋白酶组织蛋白酶 B 的正向调节。首先，我们初步数据表明肾 ENaC 活性和 MARCKS 蛋白表达呈阳性受昼夜节律蛋白 BMAL1 调节。其次，我们的初步数据显示alpha-1 与正常小鼠的肾脏相比，BMAL1 敲除小鼠肾脏中的抗胰蛋白酶含量增加野生型小鼠同时处死。三、我们的初步数据显示alpha-1 抗胰蛋白酶在肾脏中表达，强烈抑制组织蛋白酶 B 的活性，有助于血压调节。在这个项目中，我们将测试我们的假设肾 ENaC 和 MARCKS 之间的关联及其在心尖部的功能质膜与 α-1 抗胰蛋白酶表达呈负相关昼夜节律依赖方式。我们将进行实验来研究蛋白水解作用 ENaC 和 MARCKS 的顶膜表达、ENaC 活性、钠使用雄性和雌性 BMAL1 敲除小鼠 alpha-1 进行处理和血压测量抗胰蛋白酶敲除小鼠、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。顺利完成我们提出的该项目的研究将揭示 BMAL1 在肾脏 ENaC 和 MARCKS 的调节以及血压控制。我们的长期目标旨在更好地了解原发性高血压的发病机制可能会产生新的药物靶点和治疗方法。