High-level expression of rabbit 15-lipoxygenase induces collapse of the mitochondrial pH gradient in cell culture Journal Article
| Authors: | Vijayvergiya, C.; De Angelis, D.; Walther, M.; Kühn, H.; Duvoisin, R. M.; Smith, D. H.; Wiedmann, M. |
| Article Title: | High-level expression of rabbit 15-lipoxygenase induces collapse of the mitochondrial pH gradient in cell culture |
| Abstract: | 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. |
| Keywords: | controlled study; protein expression; human cell; sequence deletion; nonhuman; animal cell; mammalia; animals; cell structure; erythropoiesis; cell line; enzyme activation; in vitro study; enzyme activity; transfection; animalia; blood; amino acid sequence; molecular sequence data; cell culture; peptide fragments; recombinant proteins; cell membrane; mutagenesis, site-directed; erythrocyte; enzyme kinetics; enzyme binding; biochemistry; mitochondria; mitochondrion; cell organelle; hydrogen-ion concentration; enzyme mechanism; cell membrane permeability; experimental model; cells; isoenzyme; rabbits; intracellular membranes; biodegradation; rabbit; ph effects; cell ph; oryctolagus cuniculus; reticulocyte; humans; human; priority journal; article; red blood cells; cellular organelles; mammalian erythroblasts; arachidonate 15 lipoxygenase; arachidonate 15-lipoxygenase |
| Journal Title: | Biochemistry |
| Volume: | 43 |
| Issue: | 48 |
| ISSN: | 0006-2960 |
| Publisher: | American Chemical Society |
| Date Published: | 2004-11-09 |
| Start Page: | 15296 |
| End Page: | 15302 |
| Language: | English |
| DOI: | 10.1021/bi048745v |
| PROVIDER: | scopus |
| PUBMED: | 15568822 |
| DOI/URL: | |
| Notes: | Biochemistry -- Cited By (since 1996):7 -- Export Date: 16 June 2014 -- CODEN: BICHA -- Source: Scopus |
