Heterologous expression of a plant arginine decarboxylase gene in Trypanosoma cruzi.
Wild-type Trypanosoma cruzi epimastigotes lack arginine decarboxylase (ADC) enzymatic activity. However, the transformation of these parasites with a recombinant plasmid containing the oat ADC cDNA coding region gave rise to the transient heterologous expression of the enzyme, suggesting the absence of endogenous mechanisms that could inhibit the expression of a hypothetical own ADC gene or the assay used to measure its enzymatic activity. The foreign ADC enzyme expressed in the transgenic T. cruzi was characterized by identification of the products, the stoichiometry of the catalysed reaction, the specific inhibition by alpha-difluoromethylarginine (DFMA) and the study of its metabolic turnover. The half-life of the heterologous ADC activity in T. cruzi was about 150 min. Bioinformatics studies and polymerase chain reaction (PCR) analyses seem to indicate the absence of ADC-like DNA sequences in the wild-type T. cruzi genome.
Carrillo
Carolina
C
Fundación Instituto Leloir (formerly Instituto de Investigaciones Bioquímicas Fundación Campomar), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires and CONICET, A. Machado 151, 1405 Buenos Aires, Argentina.
Serra
María P
MP
Pereira
Claudio A
CA
Huber
Alejandra
A
González
Nélida S
NS
Algranati
Israel D
ID
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D018554:Avena / Q000235:genetics*; D001483:Base Sequence; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D014349:Trypanosoma cruzi / Q000201:enzymology / Q000235:genetics
D017931:DNA Primers; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D002262:Carboxy-Lyases; C019722:arginine decarboxylase