泛素-蛋白酶体系统抑制剂拯救细胞水平和致病性pdrin (SLC26A4)蛋白变异的离子转运功能。

IF 5.7 3区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
International journal of molecular medicine Pub Date : 2025-05-01 Epub Date: 2025-03-07 DOI:10.3892/ijmm.2025.5510
Emanuele Bernardinelli, Rapolas Jamontas, Arnoldas Matulevičius, Florian Huber, Houssein Nasser, Sophie Klaus, Haixia Zhu, Jiangang Gao, Silvia Dossena
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引用次数: 0

摘要

Pendrin (SLC26A4)是一种阴离子交换剂,在内耳、肾脏和甲状腺中大量表达,由于基因突变导致其功能障碍导致Pendred综合征和非综合征性耳聋DFNB4。pendrin蛋白的致病性变异比野生型表达少,但这种现象的机制尚不清楚。在本研究中,探讨了蛋白质表达减少源于蛋白质降解增加的假设。为了验证这一假设,在暴露于泛素-蛋白酶体系统(UPS)和溶酶体/自噬体途径抑制剂后,测量了几种致病性pendrin变体的蛋白质水平和阴离子运输功能。western blotting和定量成像检测蛋白水平;离子输运用荧光法测定。采用免疫沉淀和质谱法研究pendrin的翻译后修饰。结果显示,与野生型相比,在基于细胞的检测和Pendred综合征/DFNB4小鼠模型中,致病penddrin变体的蛋白质丰度和半衰期显著降低,表明蛋白质降解加速,而不是蛋白质产生缺陷。野生型pendrin及其变体具有丰富但不同的泛素化,这与它们不同的蛋白质稳定性是一致的。C端泛素化控制野生型penddrin的稳定性,而致病性penddrin变体p.R409H则优先发生赖氨酸77的泛素化。用研究性(MG132)或临床(硼替佐米、德兰佐米或卡非佐米)蛋白酶体抑制剂抑制UPS可挽救致病性pendrin变异的表达、质膜靶向和离子转运功能,而对溶酶体/自噬体途径的抑制无效。在所测试的化合物中,卡非佐米使pdrin p.R409H的离子转运恢复到野生型水平。这些发现表明,靶向UPS中的特定分子可以挽救penddrin蛋白致病性变体的表达和活性,这代表了Pendred综合征/DFNB4的一种新的治疗概念。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Inhibitors of the ubiquitin‑proteasome system rescue cellular levels and ion transport function of pathogenic pendrin (SLC26A4) protein variants.

Pendrin (SLC26A4) is an anion exchanger abundantly expressed in the inner ear, kidney and thyroid, and its malfunction resulting from genetic mutation leads to Pendred syndrome and non‑syndromic deafness DFNB4. Pathogenic variants of the pendrin protein are less expressed than the wild‑type, but the mechanism underlying this phenomenon is unknown. In the present study, the hypothesis that reduced protein expression stems from increased protein degradation was explored. To verify this hypothesis, the protein levels and anion transport function of several pathogenic pendrin variants were measured following exposure to inhibitors of the ubiquitin‑proteasome system (UPS) and the lysosomal/autophagosomal pathways. Protein levels were measured by western blotting and quantitative imaging; ion transport was measured with a fluorometric method. Post‑translational modification of pendrin was investigated by immunoprecipitation and mass spectrometry. The results showed that the protein abundance and half‑life of pathogenic pendrin variants were significantly reduced compared with the wild‑type in cell‑based assays and in a mouse model of Pendred syndrome/DFNB4, pointing to accelerated protein degradation rather than defective protein production. Wild‑type pendrin and its variants are abundantly but differentially ubiquitinated, consistent with their different protein stability. While ubiquitination at the C‑terminus controls the stability of wild‑type pendrin, preferential ubiquitination of lysine 77 occurred in the pathogenic pendrin variant p.R409H. Inhibition of the UPS with investigational (MG132) or clinical (bortezomib, delanzomib, or carfilzomib) proteasome inhibitors rescued the expression, plasma membrane targeting, and ion transport function of pathogenic pendrin variants, while inhibition of the lysosomal/autophagosomal pathway was ineffective. Among the compounds tested, carfilzomib rescued the ion transport of pendrin p.R409H to wild‑type levels. These findings suggest that targeting specific molecular players within the UPS can rescue the expression and activity of pathogenic variants of the pendrin protein, which represents a novel therapeutic concept for Pendred syndrome/DFNB4.

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来源期刊
International journal of molecular medicine
International journal of molecular medicine 医学-医学:研究与实验
CiteScore
12.30
自引率
0.00%
发文量
124
审稿时长
3 months
期刊介绍: The main aim of Spandidos Publications is to facilitate scientific communication in a clear, concise and objective manner, while striving to provide prompt publication of original works of high quality. The journals largely concentrate on molecular and experimental medicine, oncology, clinical and experimental cancer treatment and biomedical research. All journals published by Spandidos Publications Ltd. maintain the highest standards of quality, and the members of their Editorial Boards are world-renowned scientists.
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