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Nucleotide binding is the critical regulator of ABCG2 conformational transitions

ABCG2 is an exporter-type ABC protein that can expel numerous chemically unrelated xeno- and endobiotics from cells. When expressed in tumor cells or tumor stem cells, ABCG2 confers multidrug resistance, contributing to the failure of chemotherapy. Molecular details orchestrating substrate transloca...

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Autores principales: Gyöngy, Zsuzsanna, Mocsár, Gábor, Hegedűs, Éva, Stockner, Thomas, Ritter, Zsuzsanna, Homolya, László, Schamberger, Anita, Orbán, Tamás I, Remenyik, Judit, Szakacs, Gergely, Goda, Katalin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917445/
https://www.ncbi.nlm.nih.gov/pubmed/36763413
http://dx.doi.org/10.7554/eLife.83976
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author Gyöngy, Zsuzsanna
Mocsár, Gábor
Hegedűs, Éva
Stockner, Thomas
Ritter, Zsuzsanna
Homolya, László
Schamberger, Anita
Orbán, Tamás I
Remenyik, Judit
Szakacs, Gergely
Goda, Katalin
author_facet Gyöngy, Zsuzsanna
Mocsár, Gábor
Hegedűs, Éva
Stockner, Thomas
Ritter, Zsuzsanna
Homolya, László
Schamberger, Anita
Orbán, Tamás I
Remenyik, Judit
Szakacs, Gergely
Goda, Katalin
author_sort Gyöngy, Zsuzsanna
collection PubMed
description ABCG2 is an exporter-type ABC protein that can expel numerous chemically unrelated xeno- and endobiotics from cells. When expressed in tumor cells or tumor stem cells, ABCG2 confers multidrug resistance, contributing to the failure of chemotherapy. Molecular details orchestrating substrate translocation and ATP hydrolysis remain elusive. Here, we present methods to concomitantly investigate substrate and nucleotide binding by ABCG2 in cells. Using the conformation-sensitive antibody 5D3, we show that the switch from the inward-facing (IF) to the outward-facing (OF) conformation of ABCG2 is induced by nucleotide binding. IF-OF transition is facilitated by substrates, and hindered by the inhibitor Ko143. Direct measurements of 5D3 and substrate binding to ABCG2 indicate that the high-to-low affinity switch of the drug binding site coincides with the transition from the IF to the OF conformation. Low substrate binding persists in the post-hydrolysis state, supporting that dissociation of the ATP hydrolysis products is required to reset the high substrate affinity IF conformation of ABCG2.
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spelling pubmed-99174452023-02-11 Nucleotide binding is the critical regulator of ABCG2 conformational transitions Gyöngy, Zsuzsanna Mocsár, Gábor Hegedűs, Éva Stockner, Thomas Ritter, Zsuzsanna Homolya, László Schamberger, Anita Orbán, Tamás I Remenyik, Judit Szakacs, Gergely Goda, Katalin eLife Cell Biology ABCG2 is an exporter-type ABC protein that can expel numerous chemically unrelated xeno- and endobiotics from cells. When expressed in tumor cells or tumor stem cells, ABCG2 confers multidrug resistance, contributing to the failure of chemotherapy. Molecular details orchestrating substrate translocation and ATP hydrolysis remain elusive. Here, we present methods to concomitantly investigate substrate and nucleotide binding by ABCG2 in cells. Using the conformation-sensitive antibody 5D3, we show that the switch from the inward-facing (IF) to the outward-facing (OF) conformation of ABCG2 is induced by nucleotide binding. IF-OF transition is facilitated by substrates, and hindered by the inhibitor Ko143. Direct measurements of 5D3 and substrate binding to ABCG2 indicate that the high-to-low affinity switch of the drug binding site coincides with the transition from the IF to the OF conformation. Low substrate binding persists in the post-hydrolysis state, supporting that dissociation of the ATP hydrolysis products is required to reset the high substrate affinity IF conformation of ABCG2. eLife Sciences Publications, Ltd 2023-02-10 /pmc/articles/PMC9917445/ /pubmed/36763413 http://dx.doi.org/10.7554/eLife.83976 Text en © 2023, Gyöngy et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Gyöngy, Zsuzsanna
Mocsár, Gábor
Hegedűs, Éva
Stockner, Thomas
Ritter, Zsuzsanna
Homolya, László
Schamberger, Anita
Orbán, Tamás I
Remenyik, Judit
Szakacs, Gergely
Goda, Katalin
Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title_full Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title_fullStr Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title_full_unstemmed Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title_short Nucleotide binding is the critical regulator of ABCG2 conformational transitions
title_sort nucleotide binding is the critical regulator of abcg2 conformational transitions
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917445/
https://www.ncbi.nlm.nih.gov/pubmed/36763413
http://dx.doi.org/10.7554/eLife.83976
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