Tartrate-resistant phosphatase type 5 in Trypanosoma cruzi is important for resistance to oxidative stress promoted by hydrogen peroxide

André L A Dos-Santos; Claudia F Dick; Leandro R Lopes; Nathália Rocco-Machado; Humberto Muzi-Filho; Anita L Freitas-Mesquita; Lisvane Paes-Vieira; Adalberto Vieyra; José Roberto Meyer-Fernandes

doi:10.1016/j.exppara.2019.107748

Tartrate-resistant phosphatase type 5 in Trypanosoma cruzi is important for resistance to oxidative stress promoted by hydrogen peroxide

Exp Parasitol. 2019 Oct:205:107748. doi: 10.1016/j.exppara.2019.107748. Epub 2019 Aug 20.

Authors

André L A Dos-Santos¹, Claudia F Dick², Leandro R Lopes³, Nathália Rocco-Machado¹, Humberto Muzi-Filho⁴, Anita L Freitas-Mesquita¹, Lisvane Paes-Vieira¹, Adalberto Vieyra⁵, José Roberto Meyer-Fernandes⁶

Affiliations

¹ Leopoldo De Meis Medical Biochemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
² Leopoldo De Meis Medical Biochemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
³ Leopoldo De Meis Medical Biochemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Paulo de Góes Microbiology Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
⁴ Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Center for Structural Biology and Bioimaging (CENABIO), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
⁵ Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Center for Structural Biology and Bioimaging (CENABIO), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Graduate Program in Translational Biomedicine, Grande Rio University, Duque de Caxias, Brazil.
⁶ Leopoldo De Meis Medical Biochemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil. Electronic address: meyer@bioqmed.ufrj.br.

PMID: 31442453
DOI: 10.1016/j.exppara.2019.107748

Abstract

Trypanosoma cruzi (the causative agent of Chagas disease) presents a complex life cycle that involves adaptations in vertebrate and invertebrate hosts. As a protozoan parasite of hematophagous insects and mammalian hosts, T. cruzi is exposed to reactive oxygen species (ROS). To investigate the functionality of T. cruzi tartrate-resistant acid phosphatase type 5 (TcACP5), we cloned, superexpressed and purified the enzyme. Purified TcACP5 exhibited a V_max and apparent K_m for pNPP hydrolysis of 7.7 ± 0.2 nmol pNP × μg^-1 × h^-1 and 169.3 ± 22.6 μM, respectively. The pH dependence was characterized by sharp maximal activity at pH 5.0, and inhibition assays demonstrated its sensitivity to acid phosphatase inhibitors. Similar activities were obtained with saturating concentrations of P-Ser and P-Thr as substrates. The enzyme metabolizes hydrogen peroxide (H₂O₂) in vitro, and parasites superexpressing this enzyme were more resistant to oxidative stress promoted by H₂O₂. Taken together, these results suggest that TcACP5 plays a central role in phosphoryl transfer and redox reactions.

Keywords: Reactive oxygen species; Ser/Thr phosphatase; T. cruzi; TcACP5.

MeSH terms

Amino Acid Sequence
Fluorescent Antibody Technique
Gene Expression Regulation, Enzymologic
Hydrogen Peroxide / metabolism
Hydrogen Peroxide / pharmacology*
Hydrogen-Ion Concentration
Microscopy, Confocal
Oxidation-Reduction
Oxidative Stress / physiology*
Substrate Specificity
Tartrate-Resistant Acid Phosphatase / antagonists & inhibitors
Tartrate-Resistant Acid Phosphatase / chemistry
Tartrate-Resistant Acid Phosphatase / metabolism*
Transfection
Trypanosoma cruzi / drug effects
Trypanosoma cruzi / enzymology*

Substances

Hydrogen Peroxide
Tartrate-Resistant Acid Phosphatase