Insights on the Quest for the Structure-Function Relationship of the Mitochondrial Pyruvate Carrier

Autor(es)

Quesnay, Jose Edwin Neciosup ^{[1, 2]} ; Pollock, Naomi L. ^[3] ; Nagampalli, Raghavendra Sashi Krishna ^{[1, 4, 5]} ; Lee, Sarah C. ^[3] ; Balakrishnan, Vijayakumar ^{[6, 7]} ; Dias, Sandra Martha Gomes ^[1] ; Moraes, Isabel ^{[8, 9]} ; Dafforn, Tim R. ^[3] ; Ambrosio, Andre Luis Berteli ^[6]

Afiliação do(s) autor(es)

^[1] Ctr Res Energy & Mat, Brazilian Biosci Natl Lab, BR-13083970 Campinas - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Postgrad Program Biosci & Technol Bioact Prod, BR-13083970 Campinas - Brazil

^[3] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands - England

^[4] Monash Univ, Biomed Discovery Inst, Infect & Immun Program, Clayton, Vic 3800 - Australia

^[5] Monash Univ, Biomed Discovery Inst, Dept Biochem & Mol Biol, Clayton, Vic 3800 - Australia

^[6] Univ Sao Paulo, Sao Carlos Inst Phys, BR-13563120 Sao Carlos - Brazil

^[7] Univ Wisconsin, Sch Med & Publ Hlth, Dept Oncol, McArdle Lab Canc Res, Madison, WI 53705 - USA

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

^[9] Diamond Light Source Ltd, Membrane Prot Lab, Harwell Sci & Innovat Campus, Didcot OX11 0DE, Oxon - England

• Tipo de documento: Artigo de Revisão
• Fonte: BIOLOGY-BASEL; v. 9, n. 11 NOV 2020.
• Citações Web of Science: 0

Resumo

Simple Summary The atomic structure of a biological macromolecule determines its function. Discovering how one or more amino acid chains fold and interact to form a protein complex is critical, from understanding the most fundamental cellular processes to developing new therapies. However, this is far from a straightforward task, especially when studying a membrane protein. The functional link between the oligomeric state and complex composition of the proteins involved in the active mitochondrial transport of cytosolic pyruvate is a decades-old question but remains urgent. We present a brief historical review beginning with the identification of the so-called mitochondrial pyruvate carrier (MPC) proteins, followed by a rigorous conceptual analysis of technical approaches in more recent biochemical studies that seek to isolate and reconstitute the functional MPC complex(es) in vitro. We correlate these studies with early kinetic observations and current experimental and computational knowledge to assess their main contributions, identify gaps, resolve ambiguities, and better define the research goal. The molecular identity of the mitochondrial pyruvate carrier (MPC) was presented in 2012, forty years after the active transport of cytosolic pyruvate into the mitochondrial matrix was first demonstrated. An impressive amount of in vivo and in vitro studies has since revealed an unexpected interplay between one, two, or even three protein subunits defining different functional MPC assemblies in a metabolic-specific context. These have clear implications in cell homeostasis and disease, and on the development of future therapies. Despite intensive efforts by different research groups using state-of-the-art computational tools and experimental techniques, MPCs' structure-based mechanism remains elusive. Here, we review the current state of knowledge concerning MPCs' molecular structures by examining both earlier and recent studies and presenting novel data to identify the regulatory, structural, and core transport activities to each of the known MPC subunits. We also discuss the potential application of cryogenic electron microscopy (cryo-EM) studies of MPC reconstituted into nanodiscs of synthetic copolymers for solving human MPC2. (AU)

• Processo FAPESP: 14/20673-2 - Estudos biofísicos e bioquímicos do complexo carreador de piruvato mitocondrial e da enzima glutaminase em complexo com novos parceiros
• Beneficiário: Andre Luis Berteli Ambrosio
• Modalidade de apoio: Auxílio à Pesquisa - Regular
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• Modalidade de apoio: Auxílio à Pesquisa - Regular
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• Beneficiário: José Edwin Neciosup Quesñay
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado Direto
• Processo FAPESP: 13/07600-3 - CIBFar - Centro de Inovação em Biodiversidade e Fármacos
• Beneficiário: Glaucius Oliva
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 14/06954-9 - Estudos biofísicos, incluindo determinação da estrutura cristalográfica, do complexo Carreador de Piruvato Mitocondrial (MPC)
• Beneficiário: Raghavendra Sashi Krishna Nagampalli
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/00492-4 - Determinação da estrutura molecular da subunidade 2 do carreador mitocondrial de piruvato humano, MPC2
• Beneficiário: Vijayakumar Balakrishnan
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/02391-8 - Determinação da estrutura tridimensional de MPC2 humano por cristalografia de proteínas - uma visão mecanística do transporte mitocondrial de piruvato
• Beneficiário: Raghavendra Sashi Krishna Nagampalli
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 15/02734-7 - Estudos funcionais e estruturais de quatro homólogos recombinantes do complexo carreador mitocondrial de piruvato
• Beneficiário: José Edwin Neciosup Quesñay
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto