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The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake

The sodium taurocholate cotransporting polypeptide (NTCP) is expressed at the basolateral membrane of hepatocytes, where it mediates the uptake of conjugated bile acids and forms the hepatocyte entry receptor for the hepatitis B and D virus. Here, we aimed to identify novel protein–protein interacti...

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Autores principales: Appelman, Monique D., Robin, Marion J.D., Vogels, Esther W.M., Wolzak, Christie, Vos, Winnie G., Vos, Harmjan R., Van Es, Robert M., Burgering, Boudewijn M.T., Van de Graaf, Stan F.J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226988/
https://www.ncbi.nlm.nih.gov/pubmed/32316189
http://dx.doi.org/10.3390/cells9040986
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author Appelman, Monique D.
Robin, Marion J.D.
Vogels, Esther W.M.
Wolzak, Christie
Vos, Winnie G.
Vos, Harmjan R.
Van Es, Robert M.
Burgering, Boudewijn M.T.
Van de Graaf, Stan F.J.
author_facet Appelman, Monique D.
Robin, Marion J.D.
Vogels, Esther W.M.
Wolzak, Christie
Vos, Winnie G.
Vos, Harmjan R.
Van Es, Robert M.
Burgering, Boudewijn M.T.
Van de Graaf, Stan F.J.
author_sort Appelman, Monique D.
collection PubMed
description The sodium taurocholate cotransporting polypeptide (NTCP) is expressed at the basolateral membrane of hepatocytes, where it mediates the uptake of conjugated bile acids and forms the hepatocyte entry receptor for the hepatitis B and D virus. Here, we aimed to identify novel protein–protein interactions that could play a role in the regulation of NTCP. To this end, NTCP was precipitated from HA-tagged hNTCP-expressing HepG2 cells, and chloride channel CLIC-like 1 (CLCC1) and stomatin were identified as interacting proteins by mass spectrometry. Interaction was confirmed by co-immunoprecipitation. NTCP, CLCC1 and stomatin were found at the plasma membrane in lipid rafts, as demonstrated by a combination of immunofluorescence, cell surface biotinylation and isolation of detergent-resistant membranes. Neither CLCC1 overexpression nor its knockdown had an effect on NTCP function. However, both stomatin overexpression and knockdown increased NTCP-mediated taurocholate uptake while NTCP abundance at the plasma membrane was only increased in stomatin depleted cells. These findings identify stomatin as an interactor of NTCP and show that the interaction modulates bile salt transport.
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spelling pubmed-72269882020-05-18 The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake Appelman, Monique D. Robin, Marion J.D. Vogels, Esther W.M. Wolzak, Christie Vos, Winnie G. Vos, Harmjan R. Van Es, Robert M. Burgering, Boudewijn M.T. Van de Graaf, Stan F.J. Cells Article The sodium taurocholate cotransporting polypeptide (NTCP) is expressed at the basolateral membrane of hepatocytes, where it mediates the uptake of conjugated bile acids and forms the hepatocyte entry receptor for the hepatitis B and D virus. Here, we aimed to identify novel protein–protein interactions that could play a role in the regulation of NTCP. To this end, NTCP was precipitated from HA-tagged hNTCP-expressing HepG2 cells, and chloride channel CLIC-like 1 (CLCC1) and stomatin were identified as interacting proteins by mass spectrometry. Interaction was confirmed by co-immunoprecipitation. NTCP, CLCC1 and stomatin were found at the plasma membrane in lipid rafts, as demonstrated by a combination of immunofluorescence, cell surface biotinylation and isolation of detergent-resistant membranes. Neither CLCC1 overexpression nor its knockdown had an effect on NTCP function. However, both stomatin overexpression and knockdown increased NTCP-mediated taurocholate uptake while NTCP abundance at the plasma membrane was only increased in stomatin depleted cells. These findings identify stomatin as an interactor of NTCP and show that the interaction modulates bile salt transport. MDPI 2020-04-16 /pmc/articles/PMC7226988/ /pubmed/32316189 http://dx.doi.org/10.3390/cells9040986 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Appelman, Monique D.
Robin, Marion J.D.
Vogels, Esther W.M.
Wolzak, Christie
Vos, Winnie G.
Vos, Harmjan R.
Van Es, Robert M.
Burgering, Boudewijn M.T.
Van de Graaf, Stan F.J.
The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title_full The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title_fullStr The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title_full_unstemmed The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title_short The Lipid Raft Component Stomatin Interacts with the Na(+) Taurocholate Cotransporting Polypeptide (NTCP) and Modulates Bile Salt Uptake
title_sort lipid raft component stomatin interacts with the na(+) taurocholate cotransporting polypeptide (ntcp) and modulates bile salt uptake
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226988/
https://www.ncbi.nlm.nih.gov/pubmed/32316189
http://dx.doi.org/10.3390/cells9040986
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