Preparative enzymatic synthesis of trypanothione and trypanothione analogues.
Trypanosomatids, the causative agents of several tropical diseases, have a unique thiol metabolism based on trypanothione [bis(glutathionyl)spermidine]. Enzymes of the pathway are attractive drug target molecules but the availability of trypanothione remains an obstacle. Here, we present a convenient method for the production of trypanothione and trypanothione disulfide in >200mg quantities using a mutant of Crithidia fasciculata trypanothione synthetase in which Cys59 has been replaced by an alanine residue. The reagent costs less than 1% of the commercial price of trypanothione disulfide. The protocol also allows the synthesis of related glutathione conjugates. It will greatly facilitate the thorough analysis of this parasite's metabolism and drug screening approaches against trypanothione-dependent enzymes.
Comini
Marcelo A
MA
Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 504, 69120 Heidelberg, Germany.
Dirdjaja
Natalie
N
Kaschel
Mariel
M
Krauth-Siegel
R Luise
RL
D019732:Amide Synthases / Q000235:genetics*; D016831:Crithidia fasciculata / Q000201:enzymology*; D004220:Disulfides / Q000378:metabolism; D015195:Drug Design; D005978:Glutathione / Q000031:analogs & derivatives* / Q000096:biosynthesis; D008969:Molecular Sequence Data; D013095:Spermidine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D013379:Substrate Specificity; D014344:Trypanocidal Agents / Q000378:metabolism*; D014345:Trypanosoma / Q000201:enzymology*
D004220:Disulfides; D014344:Trypanocidal Agents; C044809:trypanothione; D019732:Amide Synthases; C064407:trypanothione synthetase; D005978:Glutathione; D013095:Spermidine