Identification of Chemical and Pharmacological Chaperones for Correction of Trafficking-Deficient Mutant Cyclic Nucleotide-Gated A3 Channels.
Identification of Chemical and Pharmacological Chaperones for Correction of Trafficking-Deficient Mutant Cyclic Nucleotide-Gated A3 Channels.
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DOI：
10.1124/molpharm.120.000180
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发表时间：
2021-06-01
影响因子：
3.6
通讯作者：
Reuter, Peggy
中科院分区：
文献类型：
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作者：
Tager, Joachim;Wissinger, Bernd;Reuter, Peggy
由错义突变引起的运输缺陷是发生在突变的、错误折叠的蛋白质中的众所周知的现象。通常，错误折叠的蛋白质被蛋白质质量控制系统保留，并被内质网相关的蛋白质降解途径降解，因此不能到达其目的地，尽管蛋白质的残余功能可以被保留。化学和药理学分子伴侣可以改善运输缺陷蛋白的靶向，因此可能是有希望的治疗应用的候选者。在这里，我们报告的应用程序的基础上的生物发光钙报告水母发光蛋白的细胞生物测定，以量化与常染色体receptor遗传性视网膜疾病色盲的突变CNGA 3通道的表面表达。77种化合物的筛选使得能够鉴定有效的化学和药理学伴侣，其导致突变CNGA 3的表面表达增加1.5至4.8倍。使用选定的化合物，我们证实了对缺陷性贩运的拯救不限于CNGA 3中的单个突变。活性化合物和我们的结构-活性相关的二氢吡啶类化合物的数据可能会提供有价值的信息，开发治疗的运输缺陷，色盲。重要性声明：本研究描述了一种新的基于发光的检测方法，以检测基于钙敏感发光蛋白水母发光蛋白的突变体运输缺陷CNGA 3通道的表面表达。使用这种方法进行化合物筛选，本研究确定了新的化学和药理学伴侣，恢复突变体贩运缺陷CNGA 3通道的表面定位。这项工作的结果可以作为开发有效化合物的起点，这些化合物可以挽救常染色体隐性遗传性视网膜疾病全色盲的运输缺陷。
Trafficking deficiency caused by missense mutations is a well known phenomenon that occurs for mutant, misfolded proteins. Typically, the misfolded protein is retained by the protein quality-control system and degraded by the endoplasmic reticulum-associated protein degradation pathway and thus does not reach its destination, although residual function of the protein may be preserved. Chemical and pharmacological chaperones can improve the targeting of trafficking-deficient proteins and thus may be promising candidates for therapeutic applications. Here, we report the application of a cellular bioassay based on the bioluminescent calcium reporter aequorin to quantify surface expression of mutant CNGA3 channels associated with the autosomal recessively inherited retinal disease achromatopsia. A screening of 77 compounds enabled the identification of effective chemical and pharmacological chaperones that result in a 1.5- to 4.8-fold increase of surface expression of mutant CNGA3. Using selected compounds, we confirmed that the rescue of the defective trafficking is not limited to a single mutation in CNGA3. Active compounds and our structure-activity correlated data for the dihydropyridine compound class may provide valuable information for developing a treatment of the trafficking defect in achromatopsia. SIGNIFICANCE STATEMENT: This study describes a novel luminescence-based assay to detect the surface expression of mutant trafficking-deficient CNGA3 channels based on the calcium-sensitive photoprotein aequorin. Using this assay for a compound screening, this study identifies novel chemical and pharmacological chaperones that restore the surface localization of mutant trafficking-deficient CNGA3 channels. The results from this work may serve as starting point for the development of potent compounds that rescue trafficking deficiencies in the autosomal recessively inherited retinal disease achromatopsia.