Probing the Ubiquitination-Mediated Function of Epithelial Sodium Channel in A549 Cells.

IF 2.4 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Current molecular pharmacology Pub Date : 2023-01-01 DOI:10.2174/1874467215666220726112002

Waheed Shabbir, Keun-Hang Susan Yang, Afzal M Dogar, Dmytro Isaev, Murat Oz

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Abstract

Background: The activity of the amiloride-sensitive epithelial sodium channel (ENaC) in the tight epithelia of the lung is regulated by proteolytic activation and ubiquitination. Pathophysiology of lung diseases is directly related to changes in one or both of these mechanisms.

Methods: In this study, we investigated the impact of ubiquitination and cathepsin-mediated proteolytic activation mechanisms on the functional regulation of ENaC in lung cancer A549 cells using the patch-clamp technique.

Results: Our findings suggest that inhibiting the proteasome (polyubiquitination) with MG132 improves ENaC activity, whereas altering the pH of the lysosome (monoubiquitination inhibition) with NH4Cl has no effect on ENaC activity. In A549 cells, inhibition of cathepsin B (CSTB) decreased the ENaC current, open probabilities (NPo and Po), and the number of active channels.

Conclusion: These findings delineate novel modes of ENaC degradation and proteolytic activation of functional channels in A549 cells. Our findings indicate that both proteolytic activation and ubiquitination of ENaC significantly affect channel function and add new insights into the endogenous ENaC processing which might help to further understand the pathophysiology of the lung disease.

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探究A549细胞上皮钠通道泛素化介导的功能。

背景:肺致密上皮中阿米洛利敏感上皮钠通道(ENaC)的活性受蛋白水解激活和泛素化调节。肺部疾病的病理生理与上述一种或两种机制的改变直接相关。方法:采用膜片钳技术，研究泛素化和组织蛋白酶介导的蛋白水解激活机制对肺癌A549细胞ENaC功能调控的影响。结果:我们的研究结果表明，用MG132抑制蛋白酶体(多泛素化)可提高ENaC活性，而用NH4Cl改变溶酶体的pH(单泛素化抑制)对ENaC活性没有影响。在A549细胞中，抑制组织蛋白酶B (CSTB)可降低ENaC电流、开放概率(NPo和Po)以及活性通道数量。结论:这些发现描述了A549细胞ENaC降解和功能通道蛋白水解激活的新模式。我们的研究结果表明，ENaC的蛋白水解激活和泛素化都显著影响通道功能，并为内源性ENaC加工提供了新的见解，这可能有助于进一步了解肺部疾病的病理生理。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current molecular pharmacology Pharmacology, Toxicology and Pharmaceutics-Drug Discovery

CiteScore

4.90

自引率

3.70%

发文量

112

期刊介绍： Current Molecular Pharmacology aims to publish the latest developments in cellular and molecular pharmacology with a major emphasis on the mechanism of action of novel drugs under development, innovative pharmacological technologies, cell signaling, transduction pathway analysis, genomics, proteomics, and metabonomics applications to drug action. An additional focus will be the way in which normal biological function is illuminated by knowledge of the action of drugs at the cellular and molecular level. The journal publishes full-length/mini reviews, original research articles and thematic issues on molecular pharmacology. Current Molecular Pharmacology is an essential journal for every scientist who is involved in drug design and discovery, target identification, target validation, preclinical and clinical development of drugs therapeutically useful in human disease.