Top

Research Article

Split Viewer

Mol. Cells 2012; 33(3): 301-307

Published online March 31, 2012

https://doi.org/10.1007/s10059-012-2260-y

© The Korean Society for Molecular and Cellular Biology

Characterization of a Putative Thioredoxin Peroxidase Prx1 of Candida albicans

Kavitha Srinivasa1, Na-Rae Kim1, Jiwon Kim1, Minsun Kim1, Ju Yun Bae2, Woojin Jeong1, Wankee Kim3,*, and Wonja Choi1,2,*

1Division of Life and Pharmaceutical Sciences, Ewha Womans University, Seoul 120-750, Korea, 2Microbial Resources Research Center, Ewha Womans University, Seoul 120-750, Korea, 3Institute for Medical Sciences, School of Medicine, Ajou University, Suwon 442-749, Korea

Correspondence to : *Correspondence: kinesin03@hanmail.net (WK); wjchoi@ewha.ac.kr (WC)

Received: November 15, 2011; Revised: December 19, 2011; Accepted: December 19, 2011

Abstract

In this study, we characterized a putative peroxidase Prx1 of Candida albicans by: 1) demonstrating the thiore-doxin-linked peroxidase activity with purified proteins, 2) examining the sensitivity to several oxidants and the accumulation of intracellular reactive oxygen species with a null mutant (prx1?), a mutant (C69S) with a point mutation at Cys69, and a revertant, and 3) subcelluar localization. Enzymatic assays showed that Prx1 is a thioredoxin-linked peroxidase which reduces both hydrogen peroxide (H2O2) and tert-butyl hydroperoxide (t-BOOH). Compared with two other strong H2O2 scavenger mutants for TSA1 and CAT1, prx1? and C69S were less sensitive to H2O2, menadione and diamide at all concentrations tested, but were more sensitive to low concentration of t-BOOH. Intracellular reactive oxygen species accumulated in prx1? and C69S cells treated with t-BOOH but not H2O2. These results suggest that peroxidase activity of Prx1 is specified to t-BOOH in cells. In both biochemical and physiological cases, the evolutionarily conserved Cys69 was found to be essential for the function. Immunocytochemical staining revealed Prx1 is localized in the cytosol of yeast cells, but is translocated to the nucleus during the hyphal transition, though the significances of this observation are unclear. Our data suggest that PRX1 has a thioredoxin peroxidase activity reducing both t-BOOH and H2O2, but its cellular function is specified to t-BOOH.

Keywords hydrogen peroxide, peroxidase, PRX1, tert-butyl hydroperoxide, translocalization

Article

Research Article

Mol. Cells 2012; 33(3): 301-307

Published online March 31, 2012 https://doi.org/10.1007/s10059-012-2260-y

Copyright © The Korean Society for Molecular and Cellular Biology.

Characterization of a Putative Thioredoxin Peroxidase Prx1 of Candida albicans

Kavitha Srinivasa1, Na-Rae Kim1, Jiwon Kim1, Minsun Kim1, Ju Yun Bae2, Woojin Jeong1, Wankee Kim3,*, and Wonja Choi1,2,*

1Division of Life and Pharmaceutical Sciences, Ewha Womans University, Seoul 120-750, Korea, 2Microbial Resources Research Center, Ewha Womans University, Seoul 120-750, Korea, 3Institute for Medical Sciences, School of Medicine, Ajou University, Suwon 442-749, Korea

Correspondence to:*Correspondence: kinesin03@hanmail.net (WK); wjchoi@ewha.ac.kr (WC)

Received: November 15, 2011; Revised: December 19, 2011; Accepted: December 19, 2011

Abstract

In this study, we characterized a putative peroxidase Prx1 of Candida albicans by: 1) demonstrating the thiore-doxin-linked peroxidase activity with purified proteins, 2) examining the sensitivity to several oxidants and the accumulation of intracellular reactive oxygen species with a null mutant (prx1?), a mutant (C69S) with a point mutation at Cys69, and a revertant, and 3) subcelluar localization. Enzymatic assays showed that Prx1 is a thioredoxin-linked peroxidase which reduces both hydrogen peroxide (H2O2) and tert-butyl hydroperoxide (t-BOOH). Compared with two other strong H2O2 scavenger mutants for TSA1 and CAT1, prx1? and C69S were less sensitive to H2O2, menadione and diamide at all concentrations tested, but were more sensitive to low concentration of t-BOOH. Intracellular reactive oxygen species accumulated in prx1? and C69S cells treated with t-BOOH but not H2O2. These results suggest that peroxidase activity of Prx1 is specified to t-BOOH in cells. In both biochemical and physiological cases, the evolutionarily conserved Cys69 was found to be essential for the function. Immunocytochemical staining revealed Prx1 is localized in the cytosol of yeast cells, but is translocated to the nucleus during the hyphal transition, though the significances of this observation are unclear. Our data suggest that PRX1 has a thioredoxin peroxidase activity reducing both t-BOOH and H2O2, but its cellular function is specified to t-BOOH.

Keywords: hydrogen peroxide, peroxidase, PRX1, tert-butyl hydroperoxide, translocalization

Mol. Cells
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

Supplementary File

Share this article on

  • line
  • mail

Related articles in Mol. Cells

Molecules and Cells

eISSN 0219-1032
qr-code Download