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Bandara 2021 Mitochondrion

From Bioblast
Publications in the MiPMap
Bandara AB, Drake JC, James CC, Smyth JW, Brown DA (2021) Complex I protein NDUFS2 is vital for growth, ROS generation, membrane integrity, apoptosis, and mitochondrial energetics. Mitochondrion 58:160-68.

Β» PMID: 33744462 Open Access

Bandara Aloka B, Drake Joshua C, James Carissa C, Smyth James W, Brown David A (2021) Mitochondrion

Abstract: Complex I is the largest and most intricate of the protein complexes of mitochondrial electron transport chain (ETC). This L-shaped enzyme consists of a peripheral hydrophilic matrix domain and a membrane-bound orthogonal hydrophobic domain. The interfacial region between these two arms is known to be critical for binding of ubiquinone moieties and has also been shown to be the binding site of Complex I inhibitors. Knowledge on specific roles of the ETC interfacial region proteins is scarce due to lack of knockout cell lines and animal models. Here we mutated nuclear encoded NADH dehydrogenase [ubiquinone] iron-sulfur protein 2 (NDUFS2), one of three protein subunits of the interfacial region, in a human embryonic kidney cell line 293 using a CRISPR/Cas9 procedure. Disruption of NDUFS2 significantly decreased cell growth in medium, Complex I specific respiration, glycolytic capacity, ATP pool and cell-membrane integrity, but significantly increased Complex II respiration, ROS generation, apoptosis, and necrosis. Treatment with idebenone, a clinical benzoquinone currently being investigated in other indications, partially restored growth, ATP pool, and oxygen consumption of the mutant. Overall, our results suggest that NDUFS2 is vital for growth and metabolism of mammalian cells, and respiratory defects of NDUFS2 dysfunction can be partially corrected with treatment of an established mitochondrial therapeutic candidate. This is the first report to use CRISPR/Cas9 approach to construct a knockout NDUFS2 cell line and use the constructed mutant to evaluate the efficacy of a known mitochondrial therapeutic to enhance bioenergetic capacity. β€’ Keywords: ATP synthesis, Apoptosis, CRISPR/Cas9, Complex I, Electron transport chain, Glycolysis, Idebenone, Necrosis, Oxygen consumption, ROS, Respiration β€’ Bioblast editor: Reiswig R β€’ O2k-Network Lab: US VA Blacksburg Brown DA

Labels: MiParea: Respiration, Genetic knockout;overexpression, Pharmacology;toxicology 

Organism: Human  Tissue;cell: HEK  Preparation: Permeabilized cells  Enzyme: Complex I 

Coupling state: LEAK, OXPHOS, ET  Pathway: N, S  HRR: Oxygraph-2k