Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter
Abstract The active transport of glycolytic pyruvate across the inner mitochondrial membrane is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa heterotypic oligomer. Here, the recombinant production of human MPC through a co-expression strategy i...
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Nature Publishing Group
2018-02-01
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Series: | Scientific Reports |
Online Access: | https://doi.org/10.1038/s41598-018-21740-z |
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doaj-f7dc7c92e807412e8c44de0a8d8ff413 |
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Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Raghavendra Sashi Krishna Nagampalli José Edwin Neciosup Quesñay Douglas Adamoski Zeyaul Islam James Birch Heitor Gobbi Sebinelli Richard Marcel Bruno Moreira Girard Carolline Fernanda Rodrigues Ascenção Angela Maria Fala Bianca Alves Pauletti Sílvio Roberto Consonni Juliana Ferreira de Oliveira Amanda Cristina Teixeira Silva Kleber Gomes Franchini Adriana Franco Paes Leme Ariel Mariano Silber Pietro Ciancaglini Isabel Moraes Sandra Martha Gomes Dias Andre Luis Berteli Ambrosio |
spellingShingle |
Raghavendra Sashi Krishna Nagampalli José Edwin Neciosup Quesñay Douglas Adamoski Zeyaul Islam James Birch Heitor Gobbi Sebinelli Richard Marcel Bruno Moreira Girard Carolline Fernanda Rodrigues Ascenção Angela Maria Fala Bianca Alves Pauletti Sílvio Roberto Consonni Juliana Ferreira de Oliveira Amanda Cristina Teixeira Silva Kleber Gomes Franchini Adriana Franco Paes Leme Ariel Mariano Silber Pietro Ciancaglini Isabel Moraes Sandra Martha Gomes Dias Andre Luis Berteli Ambrosio Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter Scientific Reports |
author_facet |
Raghavendra Sashi Krishna Nagampalli José Edwin Neciosup Quesñay Douglas Adamoski Zeyaul Islam James Birch Heitor Gobbi Sebinelli Richard Marcel Bruno Moreira Girard Carolline Fernanda Rodrigues Ascenção Angela Maria Fala Bianca Alves Pauletti Sílvio Roberto Consonni Juliana Ferreira de Oliveira Amanda Cristina Teixeira Silva Kleber Gomes Franchini Adriana Franco Paes Leme Ariel Mariano Silber Pietro Ciancaglini Isabel Moraes Sandra Martha Gomes Dias Andre Luis Berteli Ambrosio |
author_sort |
Raghavendra Sashi Krishna Nagampalli |
title |
Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
title_short |
Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
title_full |
Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
title_fullStr |
Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
title_full_unstemmed |
Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
title_sort |
human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2018-02-01 |
description |
Abstract The active transport of glycolytic pyruvate across the inner mitochondrial membrane is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa heterotypic oligomer. Here, the recombinant production of human MPC through a co-expression strategy is first described; however, substantial complex formation was not observed, and predominantly individual subunits were purified. In contrast to MPC1, which co-purifies with a host chaperone, we demonstrated that MPC2 homo-oligomers promote efficient pyruvate transport into proteoliposomes. The derived functional requirements and kinetic features of MPC2 resemble those previously demonstrated for MPC in the literature. Distinctly, chemical inhibition of transport is observed only for a thiazolidinedione derivative. The autonomous transport role for MPC2 is validated in cells when the ectopic expression of human MPC2 in yeast lacking endogenous MPC stimulated growth and increased oxygen consumption. Multiple oligomeric species of MPC2 across mitochondrial isolates, purified protein and artificial lipid bilayers suggest functional high-order complexes. Significant changes in the secondary structure content of MPC2, as probed by synchrotron radiation circular dichroism, further supports the interaction between the protein and ligands. Our results provide the initial framework for the independent role of MPC2 in homeostasis and diseases related to dysregulated pyruvate metabolism. |
url |
https://doi.org/10.1038/s41598-018-21740-z |
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doaj-f7dc7c92e807412e8c44de0a8d8ff4132020-12-08T06:08:00ZengNature Publishing GroupScientific Reports2045-23222018-02-018111310.1038/s41598-018-21740-zHuman mitochondrial pyruvate carrier 2 as an autonomous membrane transporterRaghavendra Sashi Krishna Nagampalli0José Edwin Neciosup Quesñay1Douglas Adamoski2Zeyaul Islam3James Birch4Heitor Gobbi Sebinelli5Richard Marcel Bruno Moreira Girard6Carolline Fernanda Rodrigues Ascenção7Angela Maria Fala8Bianca Alves Pauletti9Sílvio Roberto Consonni10Juliana Ferreira de Oliveira11Amanda Cristina Teixeira Silva12Kleber Gomes Franchini13Adriana Franco Paes Leme14Ariel Mariano Silber15Pietro Ciancaglini16Isabel Moraes17Sandra Martha Gomes Dias18Andre Luis Berteli Ambrosio19Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisMembrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation CampusDepartamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São PauloLaboratory of Biochemistry of Tryps – LaBTryps, Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São PauloLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratory of Biochemistry of Tryps – LaBTryps, Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São PauloDepartamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São PauloMembrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation CampusLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisLaboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e MateriaisAbstract The active transport of glycolytic pyruvate across the inner mitochondrial membrane is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa heterotypic oligomer. Here, the recombinant production of human MPC through a co-expression strategy is first described; however, substantial complex formation was not observed, and predominantly individual subunits were purified. In contrast to MPC1, which co-purifies with a host chaperone, we demonstrated that MPC2 homo-oligomers promote efficient pyruvate transport into proteoliposomes. The derived functional requirements and kinetic features of MPC2 resemble those previously demonstrated for MPC in the literature. Distinctly, chemical inhibition of transport is observed only for a thiazolidinedione derivative. The autonomous transport role for MPC2 is validated in cells when the ectopic expression of human MPC2 in yeast lacking endogenous MPC stimulated growth and increased oxygen consumption. Multiple oligomeric species of MPC2 across mitochondrial isolates, purified protein and artificial lipid bilayers suggest functional high-order complexes. Significant changes in the secondary structure content of MPC2, as probed by synchrotron radiation circular dichroism, further supports the interaction between the protein and ligands. Our results provide the initial framework for the independent role of MPC2 in homeostasis and diseases related to dysregulated pyruvate metabolism.https://doi.org/10.1038/s41598-018-21740-z |