MOLECULAR PHARMACEUTICS
·
2024
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IF 4.5
Wanjun Han,Zheyue Bo,Ting Liang,Han Liu,Lanjing Li,Zhening Guo,Ru Huan,Bruno Hagenbuch,Chunshan Gui
Organic anion-transporting polypeptides (OATPs) 1B1 and 1B3 are two highly homologous transport proteins. However, OATP1B1- and 1B3-mediated estradiol-17β-glucuronide (E17βG) uptake can be differentially affected by clotrimazole. In this study, by functional characterization on chimeric transporters and single mutants, we find that G45 in transmembrane domain 1 (TM1) and V386 in TM8 are critical for the activation of OATP1B3-mediated E17βG uptake by clotrimazole. However, the effect of clotrimazole on the function of OATP1B3 is substrate-dependent as clotrimazole does not stimulate OATP1B3-mediated uptake of 4′,5′-dibromofluorescein (DBF) and rosuvastatin. In addition, clotrimazole is not transported by OATP1B3, but it can efficiently permeate the plasma membrane due to its lipophilic properties. Homology modeling and molecular docking indicate that E17βG binds in a substrate binding pocket of OATP1B3 through hydrogen bonding and hydrophobic interactions, among which its sterol scaffold forms hydrophobic contacts with V386. In addition, a flexible glycine residue at position 45 is essential for the activation of OATP1B3. Finally, clotrimazole is predicted to bind at an allosteric site, which mainly consists of hydrophobic residues located at the cytoplasmic halves of TMs 4, 5, 10, and 11.
研究领域:
药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2022
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IF 5.895
Zhongmin Wang,Ying Li,Cecilia E. Villanueva,Taotao Peng,Wanjun Han,Zheyue Bo,Hongjian Zhang,Bruno Hagenbuch,Chunshan Gui
Estrone-3-sulfate (E3S) uptake mediated by organic anion transporting polypeptide 1B3 (OATP1B3) can be activated by epigallocatechin gallate (EGCG). In this study, by using chimeric transporters and site-directed mutagenesis, we found that Val386 in transmembrane domain 8 (TM8) is essential for OATP1B3’s activation by EGCG. Kinetic studies showed that the loss of activation of 1B3-TM8 and 1B3-V386F in the presence of EGCG is due to their decreased substrate binding affinity and reduced maximal transport rate. The overall transport efficiencies of OATP1B3, 1B3-TM8, and 1B3-V386F in the absence and presence of EGCG are 8.6 ± 0.7 vs 15.9 ± 1.4 (p < 0.05), 11.2 ± 2.1 vs 2.7 ± 0.3 (p < 0.05), and 10.2 ± 1.0 vs 2.5 ± 0.3 (p < 0.05), respectively. While 1B3-V386F cannot be activated by EGCG, its transport activity for EGCG is also diminished. OATP1B3’s activation by EGCG is substrate-dependent as EGCG inhibits OATP1B3-mediated pravastatin uptake. Furthermore, the activation of OATP1B3-mediated E3S uptake by quercetin 3-O-α-l-arabinopyranosyl(1 → 2)-α-l-rhamnopyranoside is not affected by TM8 and V386F. Taken together, the activation of OATP1B3 by small molecules is substrate- and modulator-dependent, and V386 in TM8 plays a critical role in the activation of OATP1B3-mediated E3S uptake by EGCG.
研究领域:
药理学
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
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IF 7.7
Ting Liang,Han Liu,Lanjing Li,Ru Huan,Chunshan Gui
Human organic anion transporting polypeptide 1B3 (OATP1B3) and 1B1 are two liver-specific and highly homologous uptake transporters, whose structures consist of 12 transmembrane domains. The present study showed that OATP1B3 is more heavily N-glycosylated than OATP1B1 in extracellular loop 2 (EL2) and EL5. OATP1B3 has six N-glycosylation sites, namely N134, N145, N151, N445, N503, and N516, which is twice of that of OATP1B1. Single removal of individual N-glycans seems to have minimal influence on the surface expression and function of OATP1B3. However, simultaneous removal of all N-glycans will lead to OATP1B3's large retention in the endoplasmic reticulum and cellular degradation and thus significantly disrupts its surface expression. While N-glycosylation plays a crucial role in the surface expression of OATP1B3, it also has some effect on the transport function of OATP1B3 per se , which is not due to a decrease of substrate binding affinity but due to a reduced transporter's turnover number. Taken together, N-glycosylation is essential for normal surface expression and function of OATP1B3. Its disruption by some liver diseases such as NASH might alter the pharmacokinetic/pharmacodynamic properties of OATP1B3's substrate drugs.
研究领域:
药理学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
ACS Omega
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2021
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IF 3.512
Zhongmin Wang,Ying Li,Taotao Peng,Ying Su,Xiaoting Luo,Wanjun Han,Hongjian Zhang,Jianqing Ruan,Chunshan Gui
Phenolsulfonphthalein (PSP or phenol red), a sulfonphthalein dye, has been used as a diagnostic agent and a pH indicator in cell culture medium. After administered into the body, PSP is excreted into urine and bile. The urinary excretion of PSP is mediated by organic anion transporter 1/3 (OAT1/3) and multidrug resistance protein 2 (MRP2). In biliary excretion, PSP is effluxed from hepatocytes into the bile via MRP2. However, so far, the molecular mechanism for PSP transport from the blood into hepatocytes is unclear. In the present study, six human major hepatic uptake transporters expressed on the basolateral membrane of hepatocytes, namely, organic anion transporting polypeptide 1B1 (OATP1B1), OATP1B3, OATP2B1, Na+/taurocholate cotransporting polypeptide (NTCP), organic cation transporter 1 (OCT1), and OAT2, have been investigated to see whether they are involved in the hepatic uptake of PSP. An in vitro cell-based study demonstrated that PSP is a substrate for OATP1B1, OATP1B3, and OATP2B1, with OATP1B3 showing the highest transport efficiency. The Km values for OATP1B1-, OATP1B3-, and OATP2B1-mediated PSP uptake were 11.3 ± 1.5, 7.0 ± 1.5, and 5.1 ± 1.0 μM, respectively. PSP interacts with known OATP substrates/inhibitors. However, the presence of PSP in cell culture medium has no significant effect on OATP’s function. In vivo pharmacokinetic study in wild-type and Oatp1b2-knockout mice showed that Oatp1b2-knockout led to elevated plasma concentration and decreased liver accumulation of PSP. Taken together, the present study showed that in the liver, OATP1B1, OATP1B3, and OATP2B1 are involved in the uptake of PSP from the blood into hepatocytes, which, along with MRP2-mediated efflux of PSP from hepatocytes into the bile, constitute the vectorial transport of PSP from the blood to the bile and may play a critical role in the biliary excretion of PSP.
研究领域:
药理学
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肝病学
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分子生物学
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DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
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2024
·Vol. 52
·期 11
·1323-1331
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IF 4.4
Han Liu,Lanjing Li,Ting Liang,Ru Huan,Bruno Hagenbuch,Chunshan Gui
Human organic anion transporting polypeptide (OATP) 1B1 and 1B3 are two highly homologous liver-specific uptake transporters. However, 2’,7’-dichlorofluorescein (DCF) is preferably transported by OATP1B1. In the present study, the molecular mechanisms for the selective transport of DCF by OATP1B1 were investigated by constructing and characterizing an array of OATP1B1/1B3 chimeras and site-directed mutagenesis. Our results show that transmembrane domain (TM) 10 is crucial for the surface expression and function of OATP1B1, in which Q541 and L545 play the most important roles in DCF transport. Replacement of TM10 in OATP1B1 with its OATP1B3 counterpart led to OATP1B1’s complete intracellular retention. Q541 and L545 may interact with DCF directly via hydrogen bonding and hydrophobic interactions. The decrease of DCF uptake by Q541A and L545S was due to their reduced binding affinity for DCF as compared with OATP1B1. In addition, Q541 and L545 are also crucial for the transport of estradiol-17β-glucuronide (E17βG) but not for the transport of estrone-3-sulfate (E3S), indicating different interaction modes between DCF/E17βG and E3S in OATP1B1. Taken together, Q541 and L545 in TM10 are critical for OATP1B1-mediated DCF uptake, but their effect is substrate-dependent.
研究领域:
药理学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2025
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IF 4
Lanjing Li,Ru Huan,Ting Liang,Han Liu,Yao Dong,Shuang Chen,Chen Zhang,Chunshan Gui
Estradiol is the most potent and biologically active form of estrogen with glucuronide as its predominant conjugation form. Major estradiol glucuronides include estradiol-3-glucuronide (E 2 3G), estradiol-17β-glucuronide (E 2 17βG), and estradiol-3-sulfate-17β-glucuronide (E 2 3S17βG). Hepatic disposition of E 2 17βG involves basolateral uptake transporters organic anion transporting polypeptide (OATP) 1B1/1B3 and apical efflux transporters multidrug resistance-associated protein 2 (MRP2)/breast cancer resistance protein (BCRP). In addition, E 2 3G and E 2 3S17βG are substrates for MRP2/BCRP. However, whether E 2 3G and E 2 3S17βG across the basolateral membrane of hepatocytes are transporter-mediated and which transporters play the most significant roles are largely unknown. Therefore, in the present study we investigated the human hepatic uptake transporters for E 2 3G and E 2 3S17βG. Our results showed that E 2 3G and E 2 3S17βG are transported by human OATPs. Specifically, E 2 3G is a substrate for OATP1B1, 1B3, and 2B1, with K m values being of 16.0, 23.8, and 6.4 μM, respectively, and OATP2B1 possesses the highest transport efficiency. E 2 3S17βG is a substrate for OATP1B1 and 1B3, with K m values being of 0.5 and 23.7 μM, respectively, and OATP1B1 has a higher transport efficiency. At low concentrations, E 2 3G and E 2 3S17βG are selective substrates for OATP2B1 and OATP1B1, respectively. Furthermore, mutual and differential inhibition between E 2 3G/E 2 3S17βG and known OATP substrates/inhibitors has been observed. In summary, our results demonstrated that basolateral OATPs and apical MRP2/BCRP could constitute the hepatic vectorial transport of E 2 3G and E 2 3S17βG from the blood to the bile. Significance Statement This study demonstrated that OATP1B1/1B3/2B1 are hepatic uptake transporters for E 2 3G, and OATP1B1/1B3 are hepatic uptake transporters for E 2 3S17βG. At low concentrations, E 2 3G and E 2 3S17βG are selectively transported by OATP2B1 and OATP1B1, respectively. The findings in this study unveil the underlying molecular mechanisms for the hepatic disposition of estradiol glucuronides E 2 3G and E 2 3S17βG.
研究领域:
药理学
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生物化学
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分子生物学
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生理学
·
药物代谢
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DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2026
·
IF 3.6
Han Liu,Shuang Chen,Chen Zhang,Yao Dong,Lu Yang,Jiayu Jiang,Ru Huan,Chunshan Gui
Organic anion transporting polypeptide 1B1 (OATP1B1) and 1B3 are two highly homologous liver-specific uptake transporters. While they share many common substrates, OATP1B3 exhibits selective transport for some protopanaxatriol-type (PPT-type) ginsenosides such as ginsenoside Re. In this study, we successfully identified Q422 in transmembrane domain 9 (TM9) as a key residue for OATP1B3’s selectivity towards ginsenoside Re. Substitution of Q422 with bulky residues tyrosine and phenylalanine will almost totally destroy OATP1B3’s transport for ginsenoside Re, which is not due to the reduction of transporter’s surface expression but due to the reduction of its transport activity per se . Kinetic studies showed that the decreased transport activity of Q422Y was mainly due to its reduced turnover number. Molecular docking indicated that Q422Y mutation might cause steric hindrance for ginsenoside Re binding/translocation. Q422 also plays a significant role in the transport of PPT-type ginsenoside Rg1 and notoginsenoside R1. However, it only has a trivial effect on the transport of estradiol-17β-glucuronide, estrone-3-sulfate, and rosuvastatin. Taken together, Q422 is crucial for ginsenoside transport by OATP1B3, but its role is substrate-dependent.
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DOI
高影响因子
JOURNAL OF DRUG TARGETING
·
2014
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IF 2.723
Qingyong Li,Tianyu Liu,Yunchao Li,Shuyue Luo,Qiaochu Zhu,Li Zhang,Tengfei Zhao
Camptothecin-20(s)-O-glycine ester-[N-(3′α, 12′α-dihydroxy-24′-carbonyl-5′β-cholan)] (A2), 10-(3′α,12′α-dihydroxy-5′β-cholan-24′-carboxyl)-(20 s)-camptothecin (C2), and 10-O-(3-O-(3′α, 12′α-dihydroxy-24′-carbonyl-5′β-cholan)-propyl)-(20S)-camptothecin (D2) are novel camptothecin-deoxycholic acid analogues. MTT assays were performed to assess the anticancer activity of these compounds against hepatocellular carcinoma SMMC-7721, breast carcinoma MCF-7, and colorectal carcinoma HCT-116 cells. A2 had a high killing ability on SMMC-7721 cells selectively, but C2 and D2 did not exhibit selectivity with regard to SMMC-7721 killing. Uptake assays were performed in an effort to elucidate the transport mechanisms of A2 into SMMC-7721 cells. A2 increased the mRNA expression of OATP1B3 (an organic anion-transporting polypeptide) and uptake of A2 was inhibited by rifampin (inhibitor of OATP1B3), which indicated that the transporter-mediated transport of A2 was mediated by OATP1B3. In addition, according to the western blot and apoptosis assays, we found that A2 killed SMMC-7721 cells by inducing cell apoptosis mainly via an AIF (apoptosis-inducing factor) pathway and a caspase-dependent mitochondria apoptosis pathway.
研究领域:
药理学
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肿瘤学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）