Human xenomitochondrial cybrids: Cellular models of mitochondrial complex I deficiency

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Abstract

The subunits forming the mitochondrial oxidative phosphorylation system are coded by both nuclear and mitochondrial genes. Recently, we attempted to introduce mtDNA from non-human apes into a human cell line lacking mtDNA (p°), and succeeded in producing human-common chimpanzee, human-pigmy chimpanzee, and human-gorilla xenomitochondrial cybrids (HXC). Here, we present a comprehensive characterization of oxidative phosphorylation function in these cells. Mitochondrial complexes II, III, IV, and V had activities indistinguishable from parental human or non-human primate cells. In contrast, a complex I deficiency was observed in all HXC. Kinetic studies of complex I using decylubiquinone or NADH as limiting substrates showed that the V(max) was decreased in HXC by approximately 40%, and the K(m) for the NADH was significantly increased (3-fold, p < 0.001). Rotenone inhibition studies of intact cell respiration and pyruvate-malate oxidation in permeabilized cells showed that 3 nM rotenone produced a mild effect in control cells (0-10% inhibition) but produced a marked inhibition of HXC respiration (50-75%). Immunoblotting analyses of three subunits of complex I (ND1, 75 and 49 kDa) showed that their relative amounts were not significantly altered in HXC cells. These results establish HXC as cellular models of complex I deficiency in humans and underscore the importance of nuclear and mitochondrial genomes co-evolution in optimizing oxidative phosphorylation function.

Original languageEnglish
Pages (from-to)14210-14217
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number23
DOIs
StatePublished - Jun 5 1998

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Rotenone
Gorilla gorilla
Mitochondrial DNA
NAD
Genes
Pyruvic Acid
Oxidative Phosphorylation
Cells
Pan troglodytes
Oxidation
Kinetics
Substrates
Mitochondrial complex I deficiency
Cell Respiration
Mitochondrial Genome
Mitochondrial Genes
Hominidae
Immunoblotting
Primates
Respiration

ASJC Scopus subject areas

  • Biochemistry

Cite this

Human xenomitochondrial cybrids : Cellular models of mitochondrial complex I deficiency. / Barrientos, Antonio; Kenyon, Lesley; Moraes, Carlos T.

In: Journal of Biological Chemistry, Vol. 273, No. 23, 05.06.1998, p. 14210-14217.

Research output: Contribution to journalArticle

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