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The trypanosome alternative oxidase exists as a monomer in Trypanosoma brucei mitochondria

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Abstract

The bloodstream forms of African trypanosomes solely depend on trypanosome alternative oxidase (TAO), for respiration. Similar to alternative oxidases (AOXs) found in plants and fungi, TAO is a membrane-bound diiron protein. Here, we investigated if TAO exists as a dimer like plant AOXs, or as a monomer like that of fungi. We have found that TAO forms a homo-dimer on a regular SDS-PAGE in the absence of any reducing agent and exists as a monomer under reducing condition. However, TAO does not form a dimer upon treatment of mitochondria with diamide. TAO was found as a higher molecular mass complex on a Blue-native gel after solubilization with digitonin. In the detergent soluble form, TAO activity was stimulated under reducing and inhibited under oxidizing condition. However, these conditions have no effect on the TAO activity in the mitochondria. Moreover, chemical cross-linking analysis revealed that TAO could not be cross-linked when present in the mitochondria. Together, it suggests that like certain other hydrophobic membrane proteins, TAO forms a dimer or oligomer when solubilized with detergents, and the TAO-dimer is SDS-resistant. However, it exists as a monomer in Trypanosoma brucei mitochondria.

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Acknowledgements

We thank Steve Hajduk for the ISP monoclonal antibody, Ann Umbach for soybean cotyledon mitochondria and AOA monoclonal antibody. The research was supported by NIH grants 5KO1HL03839 and 3SO6GM08037-30S1. All experiments comply with the current laws of USA.

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Authors and Affiliations

  1. Department of Microbiology, School of Medicine, Meharry Medical College, Nashville, TN, 37208, USA

    Minu Chaudhuri, Lipi Saha & Shuntae Williams

  2. Vanderbilt University School of Medicine, 301 Light Hall, Nashville, TN, 37232, USA

    Robert Daniel Ott & George C. Hill

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  1. Minu Chaudhuri
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  2. Robert Daniel Ott
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  3. Lipi Saha
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  4. Shuntae Williams
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Correspondence to Minu Chaudhuri.

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Chaudhuri, M., Ott, R.D., Saha, L. et al. The trypanosome alternative oxidase exists as a monomer in Trypanosoma brucei mitochondria. Parasitol Res 96, 178–183 (2005). https://doi.org/10.1007/s00436-005-1337-3

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  • DOI: https://doi.org/10.1007/s00436-005-1337-3

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