1. Academic Validation
  2. Protein import and oxidative folding in the mitochondrial intermembrane space of intact mammalian cells

Protein import and oxidative folding in the mitochondrial intermembrane space of intact mammalian cells

  • Mol Biol Cell. 2013 Jul;24(14):2160-70. doi: 10.1091/mbc.E12-12-0862.
Manuel Fischer 1 Sebastian Horn Anouar Belkacemi Kerstin Kojer Carmelina Petrungaro Markus Habich Muna Ali Victoria Küttner Melanie Bien Frank Kauff Jörn Dengjel Johannes M Herrmann Jan Riemer
Affiliations

Affiliation

  • 1 Cellular Biochemistry, University of Kaiserslautern, 67663 Kaiserslautern, Germany.
Abstract

Oxidation of cysteine residues to disulfides drives import of many proteins into the intermembrane space of mitochondria. Recent studies in yeast unraveled the basic principles of mitochondrial protein oxidation, but the kinetics under physiological conditions is unknown. We developed assays to follow protein oxidation in living mammalian cells, which reveal that import and oxidative folding of proteins are kinetically and functionally coupled and depend on the oxidoreductase Mia40, the sulfhydryl oxidase augmenter of liver regeneration (ALR), and the intracellular glutathione pool. Kinetics of substrate oxidation depends on the amount of Mia40 and requires tightly balanced amounts of ALR. Mia40-dependent import of Cox19 in human cells depends on the inner membrane potential. Our observations reveal considerable differences in the velocities of mitochondrial import pathways: whereas preproteins with bipartite targeting sequences are imported within seconds, substrates of Mia40 remain in the cytosol for several minutes and apparently escape premature degradation and oxidation.

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