Functional characterization of ABCB4 mutations found in progressive familial intrahepatic cholestasis type 3

Hyo Jin Park, Tae Hee Kim, So Won Kim, Shin Hye Noh, Kyeong Jee Cho, Choe Choi, Eun Young Kwon, Yang Ji Choi, Heon Yung Gee, Ji Ha Choi

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Abstract

Multidrug resistance 3 (MDR3), encoded by the ATP-binding cassette, subfamily B, member 4 gene (ABCB4), localizes to the canalicular membrane of hepatocytes and translocates phosphatidylcholine from the inner leaflet to the outer leaflet of the canalicular membrane. Progressive familial intrahepatic cholestasis type 3 (PFIC3) is a rare hepatic disease caused by genetic mutations of ABCB4. In this study, we characterized 8 ABCB4 mutations found in PFIC3 patients, using in vitro molecular assays. First, we examined the transport activity of each mutant by measuring its ATPase activity using paclitaxel or phosphatidylcholine. Then, the pathogenic mechanisms by which these mutations affect MDR3 were examined through immunoblotting, cell surface biotinylation, and immunofluorescence. As a result, three ABCB4 mutants showed significantly reduced transport activity. Among these mutants, one mutation A364V, located in intracellular domains, markedly decreased MDR3 expression on the plasma membrane, while the others did not affect the expression. The expression of MDR3 on the plasma membrane and transport activity of A364V was rescued by a pharmacological chaperone, cyclosporin A. Our study provides the molecular mechanisms of ABCB4 mutations and may contribute to the understanding of PFIC3 pathogenesis and the development of a mutation-specific targeted treatment for PFIC3.

Original languageEnglish
Article number26872
JournalScientific reports
Volume6
DOIs
Publication statusPublished - 2016 Jun 3

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Adenosine Triphosphate
Multiple Drug Resistance
Mutation
Genes
Phosphatidylcholines
Cell Membrane
Biotinylation
Membranes
Rare Diseases
Paclitaxel
Immunoblotting
Cyclosporine
Fluorescent Antibody Technique
Adenosine Triphosphatases
Progressive familial intrahepatic 3 Cholestasis
Hepatocytes
Pharmacology
Liver
Therapeutics

All Science Journal Classification (ASJC) codes

  • General

Cite this

Park, H. J., Kim, T. H., Kim, S. W., Noh, S. H., Cho, K. J., Choi, C., ... Choi, J. H. (2016). Functional characterization of ABCB4 mutations found in progressive familial intrahepatic cholestasis type 3. Scientific reports, 6, [26872]. https://doi.org/10.1038/srep26872
Park, Hyo Jin ; Kim, Tae Hee ; Kim, So Won ; Noh, Shin Hye ; Cho, Kyeong Jee ; Choi, Choe ; Kwon, Eun Young ; Choi, Yang Ji ; Gee, Heon Yung ; Choi, Ji Ha. / Functional characterization of ABCB4 mutations found in progressive familial intrahepatic cholestasis type 3. In: Scientific reports. 2016 ; Vol. 6.
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abstract = "Multidrug resistance 3 (MDR3), encoded by the ATP-binding cassette, subfamily B, member 4 gene (ABCB4), localizes to the canalicular membrane of hepatocytes and translocates phosphatidylcholine from the inner leaflet to the outer leaflet of the canalicular membrane. Progressive familial intrahepatic cholestasis type 3 (PFIC3) is a rare hepatic disease caused by genetic mutations of ABCB4. In this study, we characterized 8 ABCB4 mutations found in PFIC3 patients, using in vitro molecular assays. First, we examined the transport activity of each mutant by measuring its ATPase activity using paclitaxel or phosphatidylcholine. Then, the pathogenic mechanisms by which these mutations affect MDR3 were examined through immunoblotting, cell surface biotinylation, and immunofluorescence. As a result, three ABCB4 mutants showed significantly reduced transport activity. Among these mutants, one mutation A364V, located in intracellular domains, markedly decreased MDR3 expression on the plasma membrane, while the others did not affect the expression. The expression of MDR3 on the plasma membrane and transport activity of A364V was rescued by a pharmacological chaperone, cyclosporin A. Our study provides the molecular mechanisms of ABCB4 mutations and may contribute to the understanding of PFIC3 pathogenesis and the development of a mutation-specific targeted treatment for PFIC3.",
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Park, HJ, Kim, TH, Kim, SW, Noh, SH, Cho, KJ, Choi, C, Kwon, EY, Choi, YJ, Gee, HY & Choi, JH 2016, 'Functional characterization of ABCB4 mutations found in progressive familial intrahepatic cholestasis type 3', Scientific reports, vol. 6, 26872. https://doi.org/10.1038/srep26872

Functional characterization of ABCB4 mutations found in progressive familial intrahepatic cholestasis type 3. / Park, Hyo Jin; Kim, Tae Hee; Kim, So Won; Noh, Shin Hye; Cho, Kyeong Jee; Choi, Choe; Kwon, Eun Young; Choi, Yang Ji; Gee, Heon Yung; Choi, Ji Ha.

In: Scientific reports, Vol. 6, 26872, 03.06.2016.

Research output: Contribution to journalArticle

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AU - Kim, Tae Hee

AU - Kim, So Won

AU - Noh, Shin Hye

AU - Cho, Kyeong Jee

AU - Choi, Choe

AU - Kwon, Eun Young

AU - Choi, Yang Ji

AU - Gee, Heon Yung

AU - Choi, Ji Ha

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