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Functional Characterization Reveals the Significance of Rare Coding Variations in Human Organic Anion Transporting Polypeptide 2B1 (SLCO2B1).
Molecular Pharmaceutics ( IF 4.9 ) Pub Date : 2020-09-09 , DOI: 10.1021/acs.molpharmaceut.0c00747
Jingjie Yang 1 , Zhongmin Wang 1 , Shuai Liu 1 , Weipeng Wang 1 , Hongjian Zhang 1 , Chunshan Gui 1
Affiliation  

The organic anion transporting polypeptide 2B1 (OATP2B1), which is encoded by the SLCO2B1 gene, plays important roles in the absorption and disposition of its substrate drugs. Nonsynonymous variations of SLCO2B1 change its amino acid sequence and may alter its function. However, so far, very few genetic variants of SLCO2B1 have been functionally characterized. In the present study, first of all, 14 nonsynonymous single nucleotide variants (SNVs) of SLCO2B1 have been identified from the dbSNP database. Then, human embryonic kidney (HEK293) cells were employed as the expression system and functional studies were carried out for these 14 SNVs using substrates 4′,5′-dibromofluorescein (DBF), estrone-3-sulfate (E3S), atorvastatin, and rosuvastatin. Our results showed that four nonsynonymous rare variants, namely, SLCO2B1 c.332G > A (p.R111Q), c.1184C > A (p.P395H), c.1624G > A (p.V542M), and c.1998C > A (p.F666L), have great effect on the function of OATP2B1. Surface biotinylation and immunoblot analysis indicated that the variant c.1184C > A (p.P395H) almost completely disrupted OATP2B1’s expression on the plasma membrane. According to the three-dimensional structural model of OATP2B1 we developed, these four mutated residues are not located at the substrate binding region of OATP2B1. Their significant effect on the function of OATP2B1 could probably be attributed to jeopardizing OATP2B1’s surface expression as exemplified by c.1184C > A (p.P395H), altering the transporter’s overall structure and affecting its interactions with other proteins or the lipid bilayer. Taken together, our results demonstrated that rare coding variants could have a great impact on the function and expression of OATP2B1.

中文翻译:

功能表征揭示了人类有机阴离子转运多肽 2B1 (SLCO2B1) 中罕见编码变异的重要性。

SLCO 2 B 1 基因编码的有机阴离子转运多肽2B1 (OATP2B1)在其底物药物的吸收和处置中起重要作用。SLCO 2 B 1 的非同义变异会改变其氨基酸序列并可能改变其功能。然而,到目前为止,很少有SLCO 2 B 1 的遗传变异已被功能表征。在本研究中,首先,SLCO 2 B 的14 个非同义单核苷酸变体 (SNVs)1 个已从 dbSNP 数据库中识别出来。然后,采用人胚胎肾 (HEK293) 细胞作为表达系统,并使用底物 4',5'-二溴荧光素 (DBF)、雌酮-3-硫酸盐 (E3S)、阿托伐他汀和瑞舒伐他汀。我们的结果表明,四个非同义的稀有变体,即SLCO 2 B1 c.332G > A (p.R111Q)、c.1184C > A (p.P395H)、c.1624G > A (p.V542M)、c.1998C > A (p.F666L),对OATP2B1 的功能。表面生物素化和免疫印迹分析表明变体 c.1184C > A (p.P395H) 几乎完全破坏了 OATP2B1 在质膜上的表达。根据我们开发的OATP2B1的三维结构模型,这4个突变残基并不位于OATP2B1的底物结合区。它们对 OATP2B1 功能的显着影响可能归因于危及 OATP2B1 的表面表达,例如 c.1184C > A (p.P395H),改变转运蛋白的整体结构并影响其与其他蛋白质或脂质双层的相互作用。综合起来,
更新日期:2020-10-05
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