Mitochondria are indispensable for aerobic respiration, but many microbial eukaryotes have lost this function through reductive evolution. Their modified mitochondria are known as hydrogenosomes or mitosomes depending on whether or not they produce molecular hydrogen. The intestinal parasite Entamoeba histolytica contains mitosomes whose role in cellular metabolism is unclear. Only three proteins have been shown thus far to reside in these organelles: the molecular chaperones Hsp10 and Hsp60 and an unusual ADP/ATP carrier. Here we describe the isolation of E. histolytica mitosomes by cellular fractionation and density gradient centrifugation and show that the mitochondrial-type chaperone Hsp70 is also housed in Entamoeba mitosomes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cavalier-Smith, T. (1983). A 6 kingdom classification and a unified phylogeny, in Endocytobiology II (Schwemmler, W. and Schenk, H. E. A., eds.), De Gruyter, Berlin, pp. 1027–1034.
van der Giezen, M. and Tovar, J. (2005). Degenerate mitochondria. EMBO Rep. 6, 525–530.
Tovar, J., Fischer, A., and Clark, C.G. (1999). The mitosome, a novel organelle related to mitochondria in the amitochondrial parasite Entamoeba histolytica. Mol. Microbiol. 32, 1013–1021.
Bakker-Grunwald, T. and Wostmann, C. (1993). Entamoeba histolytica as a model for the primitive eukaryotic cell. Parasitol. Today 9, 27–31.
Clark, C. G. and Roger, A. J. (1995). Direct evidence for secondary loss of mitochondria in Entamoeba histolytica. Proc. Natl. Acad. Sci. USA 92, 6518–6521.
Loftus, B., Anderson, I., Davies, R., et al. (2005). The genome of the protist parasite Entamoeba histolytica. Nature 433, 865–868.
van der Giezen, M., Tovar, J., and Clark, C.G. (2005). Mitochondrion-derived organelles in protists and fungi. Int. Rev. Cytol. 244, 175–225.
van der Giezen, M., Leòn-Avila, G., and Tovar, J. (2005). Characterization of chaperonin 10 (Cpn10) from the intestinal human pathogen Entamoeba histolytica. Microbiology 151, 3107–3115.
Chan, K. W., Slotboom, D. J., Cox, S., et al. (2005). A novel ADP/ATP transporter in the mitosome of the microaerophilic human parasite Entamoeba histolytica. Curr. Biol. 15, 737–742.
Bakatselou, C., Kidgell, C., and Clark, C.G. (2000). A mitochondrial-type hsp70 gene of Entamoeba histolytica. Mol. Biochem. Parasitol. 110, 177–182.
Clark, C.G. and Diamond, L.S. (2002). Methods for cultivation of luminal parasitic protists of clinical importance. Clin. Microbiol. Rev. 15, 329–341.
van der Giezen, M., Birdsey, G. M., Horner, D. S., et al. (2003). Fungal hydrogenosomes contain mitochondrial heat-shock proteins. Mol. Biol. Evol. 20, 1051–1061.
Acknowledgments
We would like to thank Neil Sommerville for parasite cultures. Work in our laboratories is supported by grants from the BBSRC (BB/C507145) to JT and the Wellcome Trust (078566/A/05/Z) to MvdG.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
Tovar, J., Cox, S.S., Giezen, M.v. (2007). A Mitosome Purification Protocol Based on Percoll Density Gradients and Its Use in Validating the Mitosomal Nature of Entamoeba histolytica Mitochondrial Hsp70. In: van der Giezen, M. (eds) Protein Targeting Protocols. Methods in Molecular Biology™, vol 390. Humana Press. https://doi.org/10.1007/978-1-59745-466-7_11
Download citation
DOI: https://doi.org/10.1007/978-1-59745-466-7_11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-702-0
Online ISBN: 978-1-59745-466-7
eBook Packages: Springer Protocols