Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter

DOI: 10.1038/s41598-018-21740-z

Authors: Raghavendra Sashi Krishna Nagampalli (Centro Nacional de Pesquisa em Energia e Materiais) , José Edwin Neciosup Quesñay (Centro Nacional de Pesquisa em Energia e Materiais) , Douglas Adamoski (Centro Nacional de Pesquisa em Energia e Materiais) , Zeyaul Islam (Centro Nacional de Pesquisa em Energia e Materiais) , James Birch (Diamond Light Source) , Heitor Gobbi Sebinelli (Universidade de São Paulo) , Richard Marcel Bruno Moreira Girard (Universidade de São Paulo) , Carolline Fernanda Rodrigues Ascenção (Centro Nacional de Pesquisa em Energia e Materiais) , Angela Maria Fala (Centro Nacional de Pesquisa em Energia e Materiais) , Bianca Alves Pauletti (Centro Nacional de Pesquisa em Energia e Materiais) , Sílvio Roberto Consonni (Centro Nacional de Pesquisa em Energia e Materiais) , Juliana Ferreira De Oliveira (Centro Nacional de Pesquisa em Energia e Materiais) , Amanda Cristina Teixeira Silva (Centro Nacional de Pesquisa em Energia e Materiais) , Kleber Gomes Franchini (Centro Nacional de Pesquisa em Energia e Materiais) , Adriana Franco Paes Leme (Centro Nacional de Pesquisa em Energia e Materiais) , Ariel Mariano Silber (Universidade de São Paulo) , Pietro Ciancaglini (Universidade de São Paulo) , Isabel Moraes (Diamond Light Source) , Sandra Martha Gomes Dias (Centro Nacional de Pesquisa em Energia e Materiais) , Andre Luis Berteli Ambrosio (Centro Nacional de Pesquisa em Energia e Materiais)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 8

State: Published (Approved)
Published: February 2018

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.

Journal Keywords: Membrane proteins; Permeation and transport

Subject Areas: Biology and Bio-materials, Medicine


Instruments: B23-Circular Dichroism

Added On: 26/02/2018 10:29

Documents:
s41598-018-21740-z.pdf

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Drug Discovery Life Sciences & Biotech

Technical Tags:

Spectroscopy Circular Dichroism (CD)