Three-dimensional structure determination of human MPC2 by X-ray crystallography - a mechanistic view into mitochondrial pyruvate transport

• Grant number: 17/02391-8
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: May 01, 2017
• End date: July 31, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Andre Luis Berteli Ambrosio
• Grantee: Raghavendra Sashi Krishna Nagampalli
• Supervisor: Maria Isabel Abreu Ambrosio de Moraes
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Institution abroad: Diamond Light Source, England
• Associated to the scholarship: 14/06954-9 - Biophysical studies including X-ray structure determination of the Mitochondrial Pyruvate Carrier (MPC) complex, BP.PD

Abstract

The active transport of glycolytic pyruvate across the human inner mitochondrial membrane (IMM) is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa oligomeric heterotypic structure. We have demonstrated that homodimeric human MPC2 can promote efficient pyruvate transport into reconstituted proteoliposomes. Importantly, the derived functional requirements and kinetic features resemble those already demonstrated for pyruvate transport in mitochondrial extracts. Our results lay the initial framework for exploring the independent role of MPC2 in homeostasis and diseases related to the dysregulation of pyruvate metabolism. However, in order to establish the molecular basis of pyruvate transport, the determination of the atomic structure of human MPC2 becomes of major importance. In this context, we propose to continue our successful collaboration with the Membrane Protein Laboratory (MPL), in order to crystallize MPC2 and collect X-ray diffraction datasets using state-of-the-art facilities and experimental approaches. The experience acquired will bring benefits back to the future endeavors of the in the field of structural biology of membrane proteins.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

KRISHNA NAGAMPALLI, RAGHAVENDRA SASHI; NECIOSUP QUESNAY, JOSE EDWIN; ADAMOSKI, DOUGLAS; ISLAM, ZEYAUL; BIRCH, JAMES; SEBINELLI, HEITOR GOBBI; BRUNO MOREIRA GIRARD, RICHARD MARCEL; RODRIGUES ASCENCAO, CAROLLINE FERNANDA; FALA, ANGELA MARIA; PAULETTI, BIANCA ALVES; et al. Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter. SCIENTIFIC REPORTS, v. 8, FEB 22 2018. (17/02391-8, 14/20673-2, 16/06034-2, 15/02734-7, 15/25832-4, 14/17820-3, 14/12663-7, 14/06954-9)

QUESNAY, JOSE EDWIN NECIOSUP; POLLOCK, NAOMI L.; NAGAMPALLI, RAGHAVENDRA SASHI KRISHNA; LEE, SARAH C.; BALAKRISHNAN, VIJAYAKUMAR; DIAS, SANDRA MARTHA GOMES; MORAES, ISABEL; DAFFORN, TIM R.; AMBROSIO, ANDRE LUIS BERTELI. Insights on the Quest for the Structure-Function Relationship of the Mitochondrial Pyruvate Carrier. BIOLOGY-BASEL, v. 9, n. 11, NOV 2020. (14/20673-2, 17/11766-5, 19/02261-2, 13/07600-3, 14/06954-9, 18/00492-4, 17/02391-8, 15/02734-7)