1887

Abstract

It is becoming increasingly clear that the so-called remnant organelles of microaerophilic unicellular eukaryotes, hydrogenosomes and mitosomes, are significantly reduced versions of mitochondria. They normally lack most of the classic mitochondrial attributes, such as an electron transport chain and a genome. While hydrogenosomes generate energy by substrate-level phosphorylation along a hydrogen-producing fermentation pathway, involving iron–sulfur-cluster-containing enzymes pyruvate : ferredoxin oxidoreductase (PFO) and hydrogenase, whether mitosomes participate in ATP synthesis is currently unknown. Both enzymes were recently described in the mitosome-bearing diplomonad , also shown to produce molecular hydrogen. As published data show that giardial PFO is a membrane-associated enzyme, it could be suspected that PFO and hydrogenase operate in the mitosome, in which case the latter would by definition be a hydrogenosome. Using antibodies against recombinant enzymes of , it was shown by Western blot analysis of subcellular fractions and by confocal immunofluorescence microscopy of whole cells that neither PFO nor hydrogenase localize to the mitosome, but are mostly found in the cytosol. The giardial mitosome is known to play a role in iron–sulfur cluster assembly and to contain chaperones Cpn60 and mtHsp70, which assist, in particular, in protein import. In mitochondria, transmembrane potential is essential for this complex process. Using MitoTracker Red and organelle-specific antibodies, transmembrane potential could be detected in the hydrogenosome, but not in the mitosome. These results provide further evidence that the mitosome is one of the most highly reduced mitochondrial homologues.

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2011-06-01
2019-12-12
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