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NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes
The mitochondrial pyruvate carrier (Mpc) plays an indispensable role in the transport of pyruvates across the mitochondrial inner membrane. Despite the two distinct homologous proteins, Mpc1 and Mpc2, were identified in 2012, there are still controversies on the basic functional units and oligomeric...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10216714/ https://www.ncbi.nlm.nih.gov/pubmed/37238591 http://dx.doi.org/10.3390/biom13050719 |
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author | Wang, Zhen Ding, Wen Ruan, Maosen Liu, Yong Yang, Jing Zhang, Huiqin Shen, Bing Wang, Junfeng Li, Yunyan |
author_facet | Wang, Zhen Ding, Wen Ruan, Maosen Liu, Yong Yang, Jing Zhang, Huiqin Shen, Bing Wang, Junfeng Li, Yunyan |
author_sort | Wang, Zhen |
collection | PubMed |
description | The mitochondrial pyruvate carrier (Mpc) plays an indispensable role in the transport of pyruvates across the mitochondrial inner membrane. Despite the two distinct homologous proteins, Mpc1 and Mpc2, were identified in 2012, there are still controversies on the basic functional units and oligomeric state of Mpc complexes. In this study, yeast Mpc1 and Mpc2 proteins were expressed in a prokaryotic heterologous system. Both homo- and hetero-dimers were successfully reconstituted in mixed detergents. Interactions among Mpc monomers were recorded utilizing paramagnetic relaxation enhancement (PRE) nuclear magnetic resonance (NMR) methods. By single-channel patch-clamp assays, we discovered that both the Mpc1–Mpc2 hetero-dimer and Mpc1 homo-dimer are able to transport K(+) ions. Furthermore, the Mpc1–Mpc2 hetero-dimer demonstrated the ability to transport pyruvates, at a rate significantly higher than that of the Mpc1 homo-dimer, indicating that it could be the basic functional unit of Mpc complexes. Our findings provide valuable insights for further structural determination and the study of the transport mechanism of Mpc complexes. |
format | Online Article Text |
id | pubmed-10216714 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-102167142023-05-27 NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes Wang, Zhen Ding, Wen Ruan, Maosen Liu, Yong Yang, Jing Zhang, Huiqin Shen, Bing Wang, Junfeng Li, Yunyan Biomolecules Article The mitochondrial pyruvate carrier (Mpc) plays an indispensable role in the transport of pyruvates across the mitochondrial inner membrane. Despite the two distinct homologous proteins, Mpc1 and Mpc2, were identified in 2012, there are still controversies on the basic functional units and oligomeric state of Mpc complexes. In this study, yeast Mpc1 and Mpc2 proteins were expressed in a prokaryotic heterologous system. Both homo- and hetero-dimers were successfully reconstituted in mixed detergents. Interactions among Mpc monomers were recorded utilizing paramagnetic relaxation enhancement (PRE) nuclear magnetic resonance (NMR) methods. By single-channel patch-clamp assays, we discovered that both the Mpc1–Mpc2 hetero-dimer and Mpc1 homo-dimer are able to transport K(+) ions. Furthermore, the Mpc1–Mpc2 hetero-dimer demonstrated the ability to transport pyruvates, at a rate significantly higher than that of the Mpc1 homo-dimer, indicating that it could be the basic functional unit of Mpc complexes. Our findings provide valuable insights for further structural determination and the study of the transport mechanism of Mpc complexes. MDPI 2023-04-22 /pmc/articles/PMC10216714/ /pubmed/37238591 http://dx.doi.org/10.3390/biom13050719 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wang, Zhen Ding, Wen Ruan, Maosen Liu, Yong Yang, Jing Zhang, Huiqin Shen, Bing Wang, Junfeng Li, Yunyan NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title | NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title_full | NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title_fullStr | NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title_full_unstemmed | NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title_short | NMR and Patch-Clamp Characterization of Yeast Mitochondrial Pyruvate Carrier Complexes |
title_sort | nmr and patch-clamp characterization of yeast mitochondrial pyruvate carrier complexes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10216714/ https://www.ncbi.nlm.nih.gov/pubmed/37238591 http://dx.doi.org/10.3390/biom13050719 |
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