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Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin

Heme biosynthesis occurs through a series of reactions that take place within the cytoplasm and mitochondria, so intermediates need to move across these cellular compartments. However, the specific membrane transport mechanisms involved in the process are not yet identified. The ATP-binding cassette...

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Autores principales: Martinez, Melissa, Fendley, Gregory A., Saxberg, Alexandra D., Zoghbi, Maria E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7703921/
https://www.ncbi.nlm.nih.gov/pubmed/33253225
http://dx.doi.org/10.1371/journal.pone.0238754
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author Martinez, Melissa
Fendley, Gregory A.
Saxberg, Alexandra D.
Zoghbi, Maria E.
author_facet Martinez, Melissa
Fendley, Gregory A.
Saxberg, Alexandra D.
Zoghbi, Maria E.
author_sort Martinez, Melissa
collection PubMed
description Heme biosynthesis occurs through a series of reactions that take place within the cytoplasm and mitochondria, so intermediates need to move across these cellular compartments. However, the specific membrane transport mechanisms involved in the process are not yet identified. The ATP-binding cassette protein ABCB10 is essential for normal heme production, as knocking down this transporter in mice is embryonically lethal and accompanied by severe anemia plus oxidative damage. The role of ABCB10 is unknown, but given its location in the inner mitochondrial membrane, it has been proposed as a candidate to export either an early heme precursor or heme. Alternatively, ABCB10 might transport a molecule important for protection against oxidative damage. To help discern between these possibilities, we decided to study the effect of heme analogs, precursors, and antioxidant peptides on purified human ABCB10. Since substrate binding increases the ATP hydrolysis rate of ABC transporters, we have determined the ability of these molecules to activate purified ABCB10 reconstituted in lipid nanodiscs using ATPase measurements. Under our experimental conditions, we found that the only heme analog increasing ABCB10 ATPase activity was Zinc-mesoporphyrin. This activation of almost seventy percent was specific for ABCB10, as the ATPase activity of a negative control bacterial ABC transporter was not affected. The activation was also observed in cysteine-less ABCB10, suggesting that Zinc-mesoporphyrin’s effect did not require binding to typical heme regulatory motifs. Furthermore, our data indicate that ABCB10 was not directly activated by neither the early heme precursor delta-aminolevulinic acid nor glutathione, downsizing their relevance as putative substrates for this transporter. Although additional studies are needed to determine the physiological substrate of ABCB10, our findings reveal Zinc-mesoporphyrin as the first tool compound to directly modulate ABCB10 activity and raise the possibility that some actions of Zinc-mesoporphyrin in cellular and animal studies could be mediated by ABCB10.
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spelling pubmed-77039212020-12-03 Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin Martinez, Melissa Fendley, Gregory A. Saxberg, Alexandra D. Zoghbi, Maria E. PLoS One Research Article Heme biosynthesis occurs through a series of reactions that take place within the cytoplasm and mitochondria, so intermediates need to move across these cellular compartments. However, the specific membrane transport mechanisms involved in the process are not yet identified. The ATP-binding cassette protein ABCB10 is essential for normal heme production, as knocking down this transporter in mice is embryonically lethal and accompanied by severe anemia plus oxidative damage. The role of ABCB10 is unknown, but given its location in the inner mitochondrial membrane, it has been proposed as a candidate to export either an early heme precursor or heme. Alternatively, ABCB10 might transport a molecule important for protection against oxidative damage. To help discern between these possibilities, we decided to study the effect of heme analogs, precursors, and antioxidant peptides on purified human ABCB10. Since substrate binding increases the ATP hydrolysis rate of ABC transporters, we have determined the ability of these molecules to activate purified ABCB10 reconstituted in lipid nanodiscs using ATPase measurements. Under our experimental conditions, we found that the only heme analog increasing ABCB10 ATPase activity was Zinc-mesoporphyrin. This activation of almost seventy percent was specific for ABCB10, as the ATPase activity of a negative control bacterial ABC transporter was not affected. The activation was also observed in cysteine-less ABCB10, suggesting that Zinc-mesoporphyrin’s effect did not require binding to typical heme regulatory motifs. Furthermore, our data indicate that ABCB10 was not directly activated by neither the early heme precursor delta-aminolevulinic acid nor glutathione, downsizing their relevance as putative substrates for this transporter. Although additional studies are needed to determine the physiological substrate of ABCB10, our findings reveal Zinc-mesoporphyrin as the first tool compound to directly modulate ABCB10 activity and raise the possibility that some actions of Zinc-mesoporphyrin in cellular and animal studies could be mediated by ABCB10. Public Library of Science 2020-11-30 /pmc/articles/PMC7703921/ /pubmed/33253225 http://dx.doi.org/10.1371/journal.pone.0238754 Text en © 2020 Martinez et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Martinez, Melissa
Fendley, Gregory A.
Saxberg, Alexandra D.
Zoghbi, Maria E.
Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title_full Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title_fullStr Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title_full_unstemmed Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title_short Stimulation of the human mitochondrial transporter ABCB10 by zinc-mesoporphrin
title_sort stimulation of the human mitochondrial transporter abcb10 by zinc-mesoporphrin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7703921/
https://www.ncbi.nlm.nih.gov/pubmed/33253225
http://dx.doi.org/10.1371/journal.pone.0238754
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