High-Level Expression of Rabbit 15-Lipoxygenase Induces Collapse of the Mitochondrial pH Gradient in Cell Culture

A critical step in the development of mammalian erythroblasts into mature red blood cells is the extrusion of the nucleus, followed by intracellular degradation of the remaining organelles. It has been hypothesized that the breakdown of cellular organelles in rabbit reticulocytes is initiated by 15-...

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Bibliographic Details
Published in:Biochemistry (Easton) 2004-12, Vol.43 (48), p.15296-15302
Main Authors: Vijayvergiya, Chetan, De Angelis, Dino, Walther, Matthias, Kühn, Hartmut, Duvoisin, Robert M, Smith, Deborah H, Wiedmann, Martin
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Arachidonate 15-Lipoxygenase - biosynthesis
Arachidonate 15-Lipoxygenase - genetics
Arachidonate 15-Lipoxygenase - physiology
Cell Line
Cell Membrane Permeability - genetics
Enzyme Activation - genetics
Humans
Hydrogen-Ion Concentration
Intracellular Membranes - enzymology
Intracellular Membranes - metabolism
Mitochondria - enzymology
Mitochondria - genetics
Mitochondria - metabolism
Molecular Sequence Data
Mutagenesis, Site-Directed
Peptide Fragments - genetics
Peptide Fragments - metabolism
Peptide Fragments - pharmacology
Rabbits
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - pharmacology
Sequence Deletion
Transfection
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Summary:A critical step in the development of mammalian erythroblasts into mature red blood cells is the extrusion of the nucleus, followed by intracellular degradation of the remaining organelles. It has been hypothesized that the breakdown of cellular organelles in rabbit reticulocytes is initiated by 15-lipoxygenase. In vitro, the purified rabbit reticulocyte 15-lipoxygenase binds and permeabilizes organellar membranes, thereby releasing the lumenal contents of the organelle. Here, we demonstrate that ectopic expression of 15-lipoxygenase leads to the collapse of the mitochondrial pH gradient in nonerythroid cells, using a novel reporter of mitochondrial pH, mito-pHluorin. No change in mitochondrial pH was observed with a mutant of 15-lipoxygenase that lacks enzymatic activity. These data demonstrate that 15-lipoxygenase is capable of disrupting the pH gradient maintained by mitochondria in living cells without additional factors specific for red blood cell development.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi048745v