Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter

Author(s):Show less -

Krishna Nagampalli, Raghavendra Sashi ^[1] ; Neciosup Quesnay, Jose Edwin ^[1] ; Adamoski, Douglas ^[1] ; Islam, Zeyaul ^[1] ; Birch, James ^{[2, 3]} ; Sebinelli, Heitor Gobbi ^[4] ; Bruno Moreira Girard, Richard Marcel ^[5] ; Rodrigues Ascencao, Carolline Fernanda ^[1] ; Fala, Angela Maria ^{[1, 6]} ; Pauletti, Bianca Alves ^[1] ; Consonni, Silvio Roberto ^{[7, 1]} ; de Oliveira, Juliana Ferreira ^[1] ; Teixeira Silva, Amanda Cristina ^[1] ; Franchini, Kleber Gomes ^[1] ; Paes Leme, Adriana Franco ^[1] ; Silber, Ariel Mariano ^[5] ; Ciancaglini, Pietro ^[4] ; Moraes, Isabel ^{[2, 3, 8]} ; Gomes Dias, Sandra Martha ^[1] ; Berteli Ambrosio, Andre Luis ^[1]

Affiliation

^[1] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias, BR-13083970 Campinas, SP - Brazil

^[2] Harwell Sci & Innovat Campus, Membrane Prot Lab, Diamond Light Source, Didcot OX11 0DE, Oxon - England

^[3] Rutherford Appleton Lab, Res Complex Harwell, Didcot OX11 0FA, Oxon - England

^[4] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Pret, Dept Quim, BR-14040901 Ribeirao Preto, SP - Brazil

^[5] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, Lab Biochem Tryps LaBTryps, BR-05508000 Sao Paulo, SP - Brazil

^[6] Univ Estadual Campinas, SGC, BR-13083886 Campinas, SP - Brazil

^[7] Univ Estadual Campinas, Inst Biol, Dept Bioquim & Biol Tecidual, BR-13083862 Campinas, SP - Brazil

^[8] Natl Phys Lab, Teddington TW11 0LW, Middx - England

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 8, FEB 22 2018.
• Web of Science Citations: 6

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. (AU)

• FAPESP's process: 17/02391-8 - Three-dimensional structure determination of human MPC2 by X-ray crystallography - a mechanistic view into mitochondrial pyruvate transport
• Grantee: Raghavendra Sashi Krishna Nagampalli
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 14/20673-2 - Biophysical and biochemical studies of the mitochondrial pyruvate carrier (MPC) complex and the glutaminase enzyme bound to novel partners
• Grantee: Andre Luis Berteli Ambrosio
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/06034-2 - The biological role of amino acids and their metabolites in Trypanosoma cruzi
• Grantee: Ariel Mariano Silber
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 15/02734-7 - Functional and structural studies of four homologous recombinants of the mitochondrial pyruvate carrier complex
• Grantee: José Edwin Neciosup Quesñay
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 15/25832-4 - Metabolic regulation of genetic and epigenetic control of gene expression
• Grantee: Sandra Martha Gomes Dias
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/17820-3 - Post-transcriptional regulation of glutaminase enzyme by HuR and its relationship with high glutaminolytic levels in tumors
• Grantee: Douglas Adamoski Meira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 14/12663-7 - Biophysical and biochemical studies of the human glutaminases in complex with novel binding partners
• Grantee: Zeyaul Islam
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/06954-9 - Biophysical studies including X-ray structure determination of the Mitochondrial Pyruvate Carrier (MPC) complex
• Grantee: Raghavendra Sashi Krishna Nagampalli
• Support Opportunities: Scholarships in Brazil - Post-Doctoral