Characterization of Plasma Membrane Localization and Phosphorylation Status of Organic Anion Transporting Polypeptide (OATP) 1B1 c.521 T>C Nonsynonymous Single-Nucleotide Polymorphism.
Characterization of Plasma Membrane Localization and Phosphorylation Status of Organic Anion Transporting Polypeptide (OATP) 1B1 c.521 T>C Nonsynonymous Single-Nucleotide Polymorphism.
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DOI：
10.1007/s11095-019-2634-3
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发表时间：
2019-05-15
影响因子：
3.7
通讯作者：
Yue W
中科院分区：
文献类型：
--
作者：
Crowe A;Zheng W;Miller J;Pahwa S;Alam K;Fung KM;Rubin E;Yin F;Ding K;Yue W
膜转运蛋白有机阴离子转运多肽（OATP）1B 1介导许多药物（例如他汀类药物）的肝脏摄取。OATP 1B 1 c.521T>C（p.V174A）多态性具有降低的转运活性。关于V174 A-OATP 1B 1是否减少质膜定位，存在明确的体外结果;在生理学相关的人肝组织中未报告此类数据。尚未探索V174 A-OATP 1B 1蛋白的其他潜在变化（如磷酸化）。当前研究表征了V174 A-OATP 1B 1在基因分型人肝组织和细胞培养物中的质膜定位，并比较了V174 A-和野生型（WT）-OATP 1B 1的磷酸化状态。通过免疫组织化学在OATP 1B 1 c.521 T>C基因分型的人肝组织（n=79）中以及通过表面生物素化和共聚焦显微镜在转运蛋白过表达的人胚肾（HEK）293和HeLa细胞中测定V174 A-和WT-OATP 1B 1的定位。分别使用32 P-正磷酸盐标记和[3 H]雌二醇-17 β-葡糖苷酸蓄积测定OATP 1B 1的磷酸化和转运。所有三种方法均证明了V174 A和WT-OATP 1B 1在人肝组织和细胞培养物中的主要质膜定位。与WT-OATP 1B 1相比，V174 A-OATP 1B 1的磷酸化显著增加，转运显著减少。我们报告了与V174 A-OATP 1B 1转运功能降低相关的磷酸化增加但膜定位未受损的新发现。
Membrane transport protein organic anion transporting polypeptide (OATP) 1B1 mediates hepatic uptake of many drugs (e.g. statins). The OATP1B1 c.521T>C (p. V174A) polymorphism has reduced transport activity. Conflicting in vitro results exist regarding whether V174A-OATP1B1 has reduced plasma membrane localization; no such data has been reported in physiologically relevant human liver tissue. Other potential changes, such as phosphorylation, of the V174A-OATP1B1 protein have not been explored. Current studies characterized the plasma membrane localization of V174A-OATP1B1 in genotyped human liver tissue and cell culture and compared the phosphorylation status of V174A- and wild-type (WT)-OATP1B1. Localization of V174A- and WT-OATP1B1 were determined in OATP1B1 c.521T>C genotyped human liver tissue (n=79) by immunohistochemistry and in transporter-overexpressing human embryonic kidney (HEK) 293 and HeLa cells by surface biotinylation and confocal microscopy. Phosphorylation and transport of OATP1B1 was determined using 32P-orthophosphate labeling and [3H]estradiol-17β-glucuronide accumulation, respectively. All three methods demonstrated predominant plasma membrane localization of both V174A- and WT-OATP1B1 in human liver tissue and in cell culture. Compared to WT-OATP1B1, the V174A-OATP1B1 has significantly increased phosphorylation and reduced transport. We report novel findings of increased phosphorylation, but not impaired membrane localization, in association with the reduced transport function of the V174A-OATP1B1.
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DOI：
10.1097/00008571-200411000-00006
发表时间：
2004-11-01
期刊：
PHARMACOGENETICS
影响因子：
--
作者：
Iwai, M;Suzuki, H;Sugiyama, Y
通讯作者：
Sugiyama, Y
用于深入分析人肝脏磷酸蛋白质组的酶辅助 RP-RPLC 方法
影响因子：
3.3
作者：
Bian, Yangyang;Song, Chunxia;Zou, Hanfa
通讯作者：
Zou, Hanfa
影响因子：
6.7
作者：
Choi, J. H.;Lee, M. G.;Park, K.
通讯作者：
Park, K.
影响因子：
6.7
作者：
Chung, JY;Cho, JY;Jang, IJ
通讯作者：
Jang, IJ
影响因子：
3.6
作者：
Hahn, MK;Mazei-Robison, MC;Blakely, RD
通讯作者：
Blakely, RD