Mitochondrial complexes in Trypanosoma brucei: a novel complex and a unique oxidoreductase complex.
African trypanosomes, early diverged eukaryotes and the agents of sleeping sickness, have several basic cellular processes that are remarkably divergent from those in their mammalian hosts. They have large mitochondria and switch between oxidative phosphorylation and glycolysis as the major pathways for energy generation during their life cycle. We report here the identification and characterization of several multiprotein mitochondrial complexes from procyclic form Trypanosoma brucei. These were identified and purified using a panel of monoclonal antibodies that were generated against a submitochondrial protein fraction and using tandem affinity purification (TAP) tag affinity chromatography and localized within the cells by immunofluorescence. Protein composition analyses by mass spectrometry revealed substantial divergence of oxidoreductase complex from that of other organisms and identified a novel complex that may have a function associated with nucleic acids. The relationship to divergent physiological processes in these pathogens is discussed.
Panigrahi
Aswini K
AK
Seattle Biomedical Research Institute, Seattle, Washington 98109, USA.
Zíková
Alena
A
Dalley
Rachel A
RA
Acestor
Nathalie
N
Ogata
Yuko
Y
Anupama
Atashi
A
Myler
Peter J
PJ
Stuart
Kenneth D
KD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000913:Antibodies, Protozoan / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D005591:Chemical Fractionation; D005455:Fluorescent Antibody Technique; D047468:Immunoprecipitation; D008928:Mitochondria / Q000201:enzymology*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000302:isolation & purification / Q000378:metabolism*; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism; D020539:Sequence Analysis, Protein; D013347:Subcellular Fractions / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000166:cytology / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D024101:Mitochondrial Proteins; D046912:Multiprotein Complexes; D015800:Protozoan Proteins; D010088:Oxidoreductases