Pajuelo-Reguera 2020 Cells

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Publications in the MiPMap
Pajuelo Reguera David, ČunΓ‘tovΓ‘ KristΓ½na, VrbackΓ½ Marek, PecinovΓ‘ Alena, HouΕ‘tΔ›k Josef, MrÑček TomΓ‘Ε‘, Pecina Petr (2020) Cytochrome c oxidase subunit 4 isoform exchange results in modulation of oxygen affinity. Cells 9:E443.

Β» PMID: 32075102 Open Access

Pajuelo Reguera David, Cunatova Kristyna, Vrbacky Marek, Pecinova Alena, Houstek Josef, Mracek Tomas, Pecina Petr (2020) Cells

Abstract: Cytochrome c oxidase (COX) is regulated through tissue-, development- or environment-controlled expression of subunit isoforms. The COX4 subunit is thought to optimize respiratory chain function according to oxygen-controlled expression of its isoforms COX4i1 and COX4i2. However, biochemical mechanisms of regulation by the two variants are only partly understood. We created an HEK293-based knock-out cellular model devoid of both isoforms (COX4i1/2 KO). Subsequent knock-in of COX4i1 or COX4i2 generated cells with exclusive expression of respective isoform. Both isoforms complemented the respiratory defect of COX4i1/2 KO. The content, composition, and incorporation of COX into supercomplexes were comparable in COX4i1- and COX4i2-expressing cells. Also, COX activity, cytochrome c affinity, and respiratory rates were undistinguishable in cells expressing either isoform. Analysis of energy metabolism and the redox state in intact cells uncovered modestly increased preference for mitochondrial ATP production, consistent with the increased NADH pool oxidation and lower ROS in COX4i2-expressing cells in normoxia. Most remarkable changes were uncovered in COX oxygen kinetics. The p50 (partial pressure of oxygen at half-maximal respiration) was increased twofold in COX4i2 versus COX4i1 cells, indicating decreased oxygen affinity of the COX4i2-containing enzyme. Our finding supports the key role of the COX4i2-containing enzyme in hypoxia-sensing pathways of energy metabolism.

β€’ Bioblast editor: Gnaiger E β€’ O2k-Network Lab: CZ Prague Houstek J

Labels: MiParea: Respiration, Genetic knockout;overexpression 

Organism: Human  Tissue;cell: HEK  Preparation: Permeabilized cells, Oxidase;biochemical oxidation 

Regulation: Oxygen kinetics  Coupling state: OXPHOS, ET  Pathway: N, S, Gp, NS, Other combinations  HRR: Oxygraph-2k 

Alert2020, CZ 

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