Kinetic analyses and inhibition studies reveal novel features in peptide deformylase 1 from Trypanosoma cruzi.
In eubacteria and eukaryotic organelles N-terminal methionine excision requires the sequential action of two activities, a peptide deformylase (PDF), which systematically removes the N-formyl group present on all nascent polypeptides and methionine aminopeptidase (MAP), which exscinds methionine specifically and depends on the previous removal of the N-formyl group. In Trypanosoma cruzi two genes encoding bacterial PDF homologues have been identified and referred to as TcPDF-1 and TcPDF-2. Here we report the biochemical characterization of a truncated soluble version of TcPDF-1 lacking the hydrophobic N-terminal domain that is active with the bacterial PDF substrate formyl-methionyl-alanyl-serine but, in contrast to other PDFs, is not inhibited by actinonin. The enzyme is strongly activated by Cu(2+) and inhibited by Ni(2+). Our results show that T. cruzi PDF exhibits unique features thus providing a new avenue for the design of potential inhibitors for use in the treatment of diseases caused by trypanosomatid parasites.
Instituto de Parasitología y Biomedicina "López Neyra", Parque Tecnológico Ciencias de la Salud, Av. del Conocimiento, s/n, Consejo Superior de Investigaciones Científicas, 18100 Armilla, Granada, Spain.
Carlos A
CA
Rodrígues-Poveda
Guiomar
G
Pérez-Moreno
Antonio E
AE
Vidal
Julio A
JA
Urbina
Dolores
D
González-Pacanowska
Luis M
LM
Ruiz-Pérez
D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry; D002614:Chelating Agents / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D003470:Culture Media; D004789:Enzyme Activation; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D006877:Hydroxamic Acids / Q000494:pharmacology; D007700:Kinetics; D063208:Methionyl Aminopeptidases; D008969:Molecular Sequence Data; D009532:Nickel / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D012995:Solubility; D013379:Substrate Specificity; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics
D002614:Chelating Agents; D003470:Culture Media; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D003300:Copper; D009532:Nickel; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D000581:Amidohydrolases; C084507:peptide deformylase; C010531:actinonin