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Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding
ATP-binding cassette (ABC) multidrug transporters are large, polytopic membrane proteins that exhibit astonishing promiscuity for their transport substrates. These transporters unidirectionally efflux thousands of structurally and functionally distinct compounds. To preclude the reentry of xenobioti...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9723933/ https://www.ncbi.nlm.nih.gov/pubmed/36370844 http://dx.doi.org/10.1016/j.jbc.2022.102689 |
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author | Alhumaidi, Maryam Nentwig, Lea-Marie Rahman, Hadiar Schmitt, Lutz Rudrow, Andrew Harris, Andrzej Dillon, Cierra Restrepo, Lucas Lamping, Erwin Arya, Nidhi Ambudkar, Suresh V. Choy, John S. Golin, John |
author_facet | Alhumaidi, Maryam Nentwig, Lea-Marie Rahman, Hadiar Schmitt, Lutz Rudrow, Andrew Harris, Andrzej Dillon, Cierra Restrepo, Lucas Lamping, Erwin Arya, Nidhi Ambudkar, Suresh V. Choy, John S. Golin, John |
author_sort | Alhumaidi, Maryam |
collection | PubMed |
description | ATP-binding cassette (ABC) multidrug transporters are large, polytopic membrane proteins that exhibit astonishing promiscuity for their transport substrates. These transporters unidirectionally efflux thousands of structurally and functionally distinct compounds. To preclude the reentry of xenobiotic molecules via the drug-binding pocket, these proteins contain a highly conserved molecular gate, essentially allowing the transporters to function as molecular diodes. However, the structure–function relationship of these conserved gates and gating regions are not well characterized. In this study, we combine recent single-molecule, cryo-EM data with genetic and biochemical analyses of residues in the gating region of the yeast multidrug transporter Pdr5, the founding member of a large group of clinically relevant asymmetric ABC efflux pumps. Unlike the symmetric ABCG2 efflux gate, the Pdr5 counterpart is highly asymmetric, with only four (instead of six) residues comprising the gate proper. However, other residues in the near vicinity are essential for the gating activity. Furthermore, we demonstrate that residues in the gate and in the gating regions have multiple functions. For example, we show that Ile-685 and Val-1372 are required not only for successful efflux but also for allosteric inhibition of Pdr5 ATPase activity. Our investigations reveal that the gating region residues of Pdr5, and possibly other ABCG transporters, play a role not only in molecular gating but also in allosteric regulation, conformational switching, and protein folding. |
format | Online Article Text |
id | pubmed-9723933 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-97239332022-12-07 Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding Alhumaidi, Maryam Nentwig, Lea-Marie Rahman, Hadiar Schmitt, Lutz Rudrow, Andrew Harris, Andrzej Dillon, Cierra Restrepo, Lucas Lamping, Erwin Arya, Nidhi Ambudkar, Suresh V. Choy, John S. Golin, John J Biol Chem Research Article ATP-binding cassette (ABC) multidrug transporters are large, polytopic membrane proteins that exhibit astonishing promiscuity for their transport substrates. These transporters unidirectionally efflux thousands of structurally and functionally distinct compounds. To preclude the reentry of xenobiotic molecules via the drug-binding pocket, these proteins contain a highly conserved molecular gate, essentially allowing the transporters to function as molecular diodes. However, the structure–function relationship of these conserved gates and gating regions are not well characterized. In this study, we combine recent single-molecule, cryo-EM data with genetic and biochemical analyses of residues in the gating region of the yeast multidrug transporter Pdr5, the founding member of a large group of clinically relevant asymmetric ABC efflux pumps. Unlike the symmetric ABCG2 efflux gate, the Pdr5 counterpart is highly asymmetric, with only four (instead of six) residues comprising the gate proper. However, other residues in the near vicinity are essential for the gating activity. Furthermore, we demonstrate that residues in the gate and in the gating regions have multiple functions. For example, we show that Ile-685 and Val-1372 are required not only for successful efflux but also for allosteric inhibition of Pdr5 ATPase activity. Our investigations reveal that the gating region residues of Pdr5, and possibly other ABCG transporters, play a role not only in molecular gating but also in allosteric regulation, conformational switching, and protein folding. American Society for Biochemistry and Molecular Biology 2022-11-10 /pmc/articles/PMC9723933/ /pubmed/36370844 http://dx.doi.org/10.1016/j.jbc.2022.102689 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Alhumaidi, Maryam Nentwig, Lea-Marie Rahman, Hadiar Schmitt, Lutz Rudrow, Andrew Harris, Andrzej Dillon, Cierra Restrepo, Lucas Lamping, Erwin Arya, Nidhi Ambudkar, Suresh V. Choy, John S. Golin, John Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title | Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title_full | Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title_fullStr | Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title_full_unstemmed | Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title_short | Residues forming the gating regions of asymmetric multidrug transporter Pdr5 also play roles in conformational switching and protein folding |
title_sort | residues forming the gating regions of asymmetric multidrug transporter pdr5 also play roles in conformational switching and protein folding |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9723933/ https://www.ncbi.nlm.nih.gov/pubmed/36370844 http://dx.doi.org/10.1016/j.jbc.2022.102689 |
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