外排转运体多药耐药蛋白4 (MRP4, ABCC4)编码变异与预测功能影响的体外表征

IF 1.7 3区 医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Jaymie Mailloux, Samantha Medwid, Amanda Facey, Inmo Sung, Laura E Russell, Rommel G Tirona, Richard B Kim, Ute I Schwarz
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引用次数: 2

摘要

MRP4(基因ABCC4)是一种多态外排转运蛋白,与药物诱导的毒性有关。我们在欧洲人中选择了10个常见的MRP4编码变异进行实验表征,其中9个变异预计是有害的或功能性的(综合注释依赖耗尽评分>15)。在HEK293T过表达MRP4野生型或变体中,通过与摄取转运体共转染优化细胞底物负载,我们通过量化细胞内两种原型底物,牛油胆酸(TCA)和雌二醇17-β-葡萄糖醛酸(E217βG)的积累来评估蛋白质的定位和活性。与MRP4野生型相比,以前未研究过的新变体V458M和T1142M对E217βG和TCA的活性降低
本文章由计算机程序翻译,如有差异,请以英文原文为准。
In-vitro characterization of coding variants with predicted functional implications in the efflux transporter multidrug resistance protein 4 (MRP4, ABCC4).

MRP4 (gene ABCC4) is a polymorphic efflux transporter that has been implicated in drug-induced toxicity. We selected ten commonly observed MRP4 coding variants among Europeans for experimental characterization including nine variants predicted to be deleterious or functional (combined annotation-dependent depletion score >15). We assessed protein localization and activity by quantifying intracellular accumulation of two prototypic substrates, taurocholic acid (TCA) and estradiol 17-β-glucuronide (E217βG), in HEK293T over-expressing MRP4 wildtype or variant where cellular substrate loading was optimized through co-transfection with an uptake transporter. V458M, a novel variant not previously studied, and T1142M, showed reduced activity compared to MRP4 wildtype for E217βG and TCA (P < 0.01), while L18I, G187W, K293E, and R531Q moderately increased activity in a substrate-dependent manner. Protein expression analysis indicated reduced cell surface expression for V458M (P < 0.01) but not T1142M compared to wildtype. Reduced activity may result from altered surface expression (V458M) or intrinsic activity as both variants map within the nucleotide-binding domains of MRP4. G187W showed a trend for reduced surface expression (P = 0.054) despite transport comparable or increased to wildtype suggesting enhanced intrinsic activity. Our findings suggest moderately altered MRP4 activity in six out of nine predicted functional variants with likely different mechanisms and substrate-specific effects. Cell-based studies using multiple known substrates are warranted to more accurately predict functional variants in this clinically important transporter.

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来源期刊
Pharmacogenetics and genomics
Pharmacogenetics and genomics 医学-生物工程与应用微生物
CiteScore
3.20
自引率
3.80%
发文量
47
审稿时长
3 months
期刊介绍: ​​​​Pharmacogenetics and Genomics is devoted to the rapid publication of research papers, brief review articles and short communications on genetic determinants in response to drugs and other chemicals in humans and animals. The Journal brings together papers from the entire spectrum of biomedical research and science, including biochemistry, bioinformatics, clinical pharmacology, clinical pharmacy, epidemiology, genetics, genomics, molecular biology, pharmacology, pharmaceutical sciences, and toxicology. Under a single cover, the Journal provides a forum for all aspects of the genetics and genomics of host response to exogenous chemicals: from the gene to the clinic.
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