Dihydrolipoamide dehydrogenase in the trypanosoma subgenus, trypanozoon.
The enzyme dihydrolipoamide dehydrogenase has been discovered and characterised in four salivarian trypanosomes of the subgenus trypanozoon: Trypanosoma brucei brucei, T. b. gambiense, T. b. rhodesiense, and Trypanosoma evansi. The three T. brucei species, which have insect procyclic forms biochemically distinct from their mammalian bloodstream forms, express dihydrolipoamide dehydrogenase in both cell types, but have higher levels in the procyclic forms. Determination of Michaelis constants for the enzyme from each of the three T. brucei species did not reveal any significant kinetic differences between the bloodstream and procyclic enzymes. On Western blots, antibodies raised against dihydrolipoamide dehydrogenase from the stereorarian trypanosome, Trypanosoma cruzi, cross-react strongly with the dihydrolipoamide dehydrogenase from all three T. brucei species; by this method, the relative molecular masses of their dihydrolipoamide dehydrogenases are indistinguishable. Dihydrolipoamide dehydrogenase was purified from both the bloodstream and the procyclic forms of T. b. brucei, and the N-terminal have been sequenced. These sequences are identical to the derived protein sequence of the cloned gene (Else et al., Eur. J. Biochem. 212 (1993) 423-429), but have a nine amino acid N-terminal truncation, giving an N-terminus equivalent to that of T. cruzi dihydrolipoamide dehydrogenase. The T. b. brucei dihydrolipoamide dehydrogenase gene has been expressed in Escherichia coli and the resultant protein purified; its N-terminus is processed in a similar fashion to that in the trypanosome, but with reduced specificity.
Else
A J
AJ
Department of Biochemistry, University of Bath, England.
Clarke
J F
JF
Willis
A
A
Jackman
S A
SA
Hough
D W
DW
Danson
M J
MJ
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan / Q000235:genetics; D003001:Cloning, Molecular; D003429:Cross Reactions; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D013045:Species Specificity; D014345:Trypanosoma / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D014352:Trypanosomiasis / Q000469:parasitology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D008058:Dihydrolipoamide Dehydrogenase