Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications
The human P-glycoprotein (P-gp), a transporter responsible for multidrug resistance, is present in the plasma membrane’s raft and non-raft domains. One specific conformation of P-gp that binds to the monoclonal antibody UIC2 is primarily associated with raft domains and displays heightened internali...
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MDPI AG
2023-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/15/12335 |
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| author | Zsuzsanna Gutay-Tóth Gabriella Gellen Minh Doan James F. Eliason János Vincze Lajos Szente Ferenc Fenyvesi Katalin Goda Miklós Vecsernyés Gábor Szabó Zsolt Bacso |
| author_facet | Zsuzsanna Gutay-Tóth Gabriella Gellen Minh Doan James F. Eliason János Vincze Lajos Szente Ferenc Fenyvesi Katalin Goda Miklós Vecsernyés Gábor Szabó Zsolt Bacso |
| author_sort | Zsuzsanna Gutay-Tóth |
| collection | DOAJ |
| description | The human P-glycoprotein (P-gp), a transporter responsible for multidrug resistance, is present in the plasma membrane’s raft and non-raft domains. One specific conformation of P-gp that binds to the monoclonal antibody UIC2 is primarily associated with raft domains and displays heightened internalization in cells overexpressing P-gp, such as in NIH-3T3 MDR1 cells. Our primary objective was to investigate whether the trafficking of this particular P-gp conformer is dependent on cholesterol levels. Surprisingly, depleting cholesterol using cyclodextrin resulted in an unexpected increase in the proportion of raft-associated P-gp within the cell membrane, as determined by UIC2-reactive P-gp. This increase appears to be a compensatory response to cholesterol loss from the plasma membrane, whereby cholesterol-rich raft micro-domains are delivered to the cell surface through an augmented exocytosis process. Furthermore, this exocytotic event is found to be part of a complex trafficking mechanism involving lysosomal exocytosis, which contributes to membrane repair after cholesterol reduction induced by cyclodextrin treatment. Notably, cells overexpressing P-gp demonstrated higher total cellular cholesterol levels, an increased abundance of stable lysosomes, and more effective membrane repair following cholesterol modifications. These modifications encompassed exocytotic events that involved the transport of P-gp-carrying rafts. Importantly, the enhanced membrane repair capability resulted in a durable phenotype for MDR1 expressing cells, as evidenced by significantly improved viabilities of multidrug-resistant Pgp-overexpressing immortal NIH-3T3 MDR1 and MDCK-MDR1 cells compared to their parents when subjected to cholesterol alterations. |
| first_indexed | 2024-03-11T00:25:30Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T00:25:30Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-741cecfa691849138d487da610888caa2023-11-18T23:03:36ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-08-0124151233510.3390/ijms241512335Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol ModificationsZsuzsanna Gutay-Tóth0Gabriella Gellen1Minh Doan2James F. Eliason3János Vincze4Lajos Szente5Ferenc Fenyvesi6Katalin Goda7Miklós Vecsernyés8Gábor Szabó9Zsolt Bacso10Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryGreat Lakes Stem Cell Innovation Center, Detroit, MI 48202, USADepartment of Physiology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryCycloLab Cyclodextrin Research & Development Laboratory, Ltd., 1097 Budapest, HungaryDepartment of Pharmaceutical Technology, Faculty of Pharmacy, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Pharmaceutical Technology, Faculty of Pharmacy, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryThe human P-glycoprotein (P-gp), a transporter responsible for multidrug resistance, is present in the plasma membrane’s raft and non-raft domains. One specific conformation of P-gp that binds to the monoclonal antibody UIC2 is primarily associated with raft domains and displays heightened internalization in cells overexpressing P-gp, such as in NIH-3T3 MDR1 cells. Our primary objective was to investigate whether the trafficking of this particular P-gp conformer is dependent on cholesterol levels. Surprisingly, depleting cholesterol using cyclodextrin resulted in an unexpected increase in the proportion of raft-associated P-gp within the cell membrane, as determined by UIC2-reactive P-gp. This increase appears to be a compensatory response to cholesterol loss from the plasma membrane, whereby cholesterol-rich raft micro-domains are delivered to the cell surface through an augmented exocytosis process. Furthermore, this exocytotic event is found to be part of a complex trafficking mechanism involving lysosomal exocytosis, which contributes to membrane repair after cholesterol reduction induced by cyclodextrin treatment. Notably, cells overexpressing P-gp demonstrated higher total cellular cholesterol levels, an increased abundance of stable lysosomes, and more effective membrane repair following cholesterol modifications. These modifications encompassed exocytotic events that involved the transport of P-gp-carrying rafts. Importantly, the enhanced membrane repair capability resulted in a durable phenotype for MDR1 expressing cells, as evidenced by significantly improved viabilities of multidrug-resistant Pgp-overexpressing immortal NIH-3T3 MDR1 and MDCK-MDR1 cells compared to their parents when subjected to cholesterol alterations.https://www.mdpi.com/1422-0067/24/15/12335ABCB1 transportercyclodextrinmembrane repairrafttraffickingUIC2 |
| spellingShingle | Zsuzsanna Gutay-Tóth Gabriella Gellen Minh Doan James F. Eliason János Vincze Lajos Szente Ferenc Fenyvesi Katalin Goda Miklós Vecsernyés Gábor Szabó Zsolt Bacso Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications International Journal of Molecular Sciences ABCB1 transporter cyclodextrin membrane repair raft trafficking UIC2 |
| title | Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications |
| title_full | Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications |
| title_fullStr | Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications |
| title_full_unstemmed | Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications |
| title_short | Cholesterol-Depletion-Induced Membrane Repair Carries a Raft Conformer of P-Glycoprotein to the Cell Surface, Indicating Enhanced Cholesterol Trafficking in MDR Cells, Which Makes Them Resistant to Cholesterol Modifications |
| title_sort | cholesterol depletion induced membrane repair carries a raft conformer of p glycoprotein to the cell surface indicating enhanced cholesterol trafficking in mdr cells which makes them resistant to cholesterol modifications |
| topic | ABCB1 transporter cyclodextrin membrane repair raft trafficking UIC2 |
| url | https://www.mdpi.com/1422-0067/24/15/12335 |
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