Abstract
Thiol-based peroxiredoxins (Prxs) are conserved throughout all kingdoms. We have found that a conserved typical 2-Cys Prx-like protein (PaPrx) from Pseudomonas aeruginosa bacteria displays diversity in its structure and apparent molecular weight (MW), and can act alternatively as a peroxidase and molecular chaperone. We have also identified a regulatory factor involved in this structural and functional switching. Exposure of P. aeruginosa to hydrogen peroxide (H2O2) causes PaPrx to convert from a high MW (HMW) complex to a low MW (LMW) form, which triggers a chaperone to peroxidase functional switch. This structural switching is primarily guided by either the thioredoxin (Trx) or glutathione (GSH) systems. Furthermore, comparison of our structural data [native and non-reducing polyacrylamide gel electrophoresis (PAGE) analysis, size exclusion chromatography (SEC) analysis, and electron microscopy (EM) observations] and enzymatic analyses (peroxidase and chaperone assay) revealed that the formation of oligomeric HMW complex structures increased chaperone activity of PaPrx. These results suggest that multimerization of PaPrx complexes promotes chaperone activity, and dissociation of the complexes into LMW species enhances peroxidase activity. Thus, the dual functions of PaPrx are clearly associated with their ability to form distinct protein structures.
Similar content being viewed by others
References
Alphey, M.S., Bond, C.S., Tetaud, E., Fairlamb, A.H., and Hunter, W.N. (2000). The structure of reduced tryparedoxin peroxidase reveals a decamer and insight into reactivity of 2 Cysperoxiredoxins. J. Mol. Biol. 300, 903–916.
Arrigo, A.P. (1998). Small stress proteins: chaperones that act as regulators of intracellular redox state and programmed cell death. Biol. Chem. 379, 19–26.
Barford, D. (2004). The role of cysteine residues as redox-sensitive regulatory switches. Curr. Opin. Struct. Biol. 14, 679–686.
Burgess, S.A., Walker, M.L., Thirumurugan, K., Trinick, J., and Knight, P.J. (2004). Use of negative stain and single-particle image processing to explore dynamic properties of flexible macromolecules. J. Struct. Biol. 147, 247–258.
Chae, H.Z., Robison, K., Poole, L.B., Church, G., Storz, G., and Rhee, S.G. (1994). Cloning and sequencing of thiol-specific antioxidant from mammalian brain: alkyl hydroperoxide reductase and thiol-specific antioxidant define a large family of antioxidant enzymes. Proc. Natl. Acad. Sci. USA 91, 7017–7021.
Chang, T.S., Jeong, W., Choi, S.Y., Yu, S., Kang, S.W., and Rhee, S.G. (2002). Regulation of peroxiredoxin I activity by Cdc2-mediated phosphorylation. J. Biol. Chem. 277, 25370–25376.
Chauhan, R., and Mande, S.C. (2001). Characterization of the Mycobacterium tuberculosis H37Rv alkyl hydroperoxidase AhpC points to the importance of ionic interactions in oligomerization and activity. Biochem. J. 354, 209–215.
Cheong, N.E., Choi, Y.O., Lee, K.O., Kim, W.Y., Jung, B.G., Chi, Y.H., Jeong, J.S., Kim, K., Cho, M.J., and Lee, S.Y. (1999). Molecular cloning, expression, and functional characterization of a 2Cys-peroxiredoxin in Chinese cabbage. Plant Mol. Biol. 40, 825–834.
Chuang, M.H., Wu, M.S., Lo, W.L., Lin, J.T., Wong, C.H., and Chiou, S.H. (2006). The antioxidant protein alkylhydroperoxide reductase of Helicobacter pylori switches from a peroxide reductase to a molecular chaperone function. Proc. Natl. Acad. Sci. USA 103, 2552–2557.
Haley, D., Horwitz, J., and Stewart, P.L. (1998). The small heat shock protein, αB-crystallin, has a variable quaternary structure. J. Mol. Biol. 277, 27–35.
Hartl, F.U. (1996). Molecular chaperone in cellular protein folding. Nature 381, 571–580.
Hendrick, J.P., and Hartl, F.U. (1993). Molecular chaperone functions of heat shock proteins. Annu. Rev. Biochem. 62, 349–384.
Hirotsu, S., Abe, Y., Okada, K., Nagahara, N., Hori, H., Nishino, T., and Hakoshima, T. (1999). Crystal structure of a multifunctional 2-Cys peroxiredoxin heme-binding protein 23 kDa/proliferation-associated gene product. Proc. Natl. Acad. Sci. USA 96, 12333–12338.
Hofmann, B., Hecht, H.J., and Flohé, L. (2002). Peroxiredoxins. Biol. Chem. 383(3–4), 347–364.
Ito, H., Kamei, K., Iwamoto, I., Inaguma, Y., Nohara, D., and Kato, K. (2001). Phosphorylation-induced change of the oligomerization state of alpha B-crystallin, J. Biol. Chem. 276, 5346–5352.
Jang, H.H., Lee, K.O., Chi, Y.H., Jung, B.G., Park, S.K., Park, J.H., Lee, J.R., Lee, S.S., Moon, J.C., Yun, J.W., et al. (2004). Two enzymes in one; two yeast peroxiredoxins display oxidative stress-dependent switching from a peroxidase to a molecular chaperone function. Cell 117, 625–635.
Jang, H.H., Chi, Y.H., Park, S.K., Lee, S.S., Lee, J.R., Park, J.H., Moon, J.C., Lee, Y.M., Kim, S.Y., Lee, K.H., et al. (2006). Structural and functional regulation of eukaryotic 2-Cys peroxiredoxins including the plant ones in cellular defense signaling mechanisms against oxidative stress. Physiol. Plant. 126, 549–559.
Jeong, W.J., Cha, M.K., and Kim, I.H. (2000). A new member of human Tsa/AhpC as thioredoxin-dependent thiol peroxidase. J. Biochem. Mol. Biol. 33, 234–241.
Kim, K.S., Choi, S.Y., Kwon, H.Y., Won, M.H., Kang, T.C., and Kang, J.H. (2002). Aggregation of α-synuclein induced by the Cu, Zn-superoxide dismutase and hydrogen peroxide system. Free Radic. Biol. Med. 32, 544–550.
Kim, J.A., Park, S., Kim, K., Rhee, S.G., and Kang, S.W. (2005). Activity assay of mammalian 2-cys peroxiredoxins using yeast thioredoxin reductase system. Anal. Biochem. 338, 216–223.
Kitano, K., Niimura, Y., Nishiyama, Y., and Miki, K. (1999). Stimulation of peroxidase activity by decamerization related to ionic strength: ahpC protein from Amphibacillus xylanus. J. Biochem. 126, 313–319.
Lee, G.J., Roseman, A.M., Saibil, H.R., and Vierling, E. (1997). A small heat shock protein stably binds heat-denatured model substrates and can maintain a substrate in a folding-competent state. EMBO J. 16, 659–671.
Lee, J.R., Lee, S.S., Jang, H.H., Lee, Y.M., Park, J.H., Park, S.-C., Moon, J.C., Park, S.K., Kim, S.Y., Lee, S.Y., et al. (2009). Heatshock dependent oligomeric status alters the function of a plantspecific thioredoxin-like protein, AtTDX. Proc. Natl. Acad. Sci. USA 106, 5978–5983.
Moon, J.C., Hah, Y.S., Kim, W.Y., Jung, B.G., Jang, H.H., Lee, J.R., Kim, S.Y., Lee, Y.M., Jeon, M.K., Kim, C.W., et al. (2005). Oxidative stress-dependent structural and functional switching of a human 2-Cys peroxiredoxin isotype II that enhances HeLa cell resistance to H2O2-induced cell death, J. Biol. Chem. 280, 28775–28784.
Nooren, I.M.A., and Thornton, J.M. (2003a). Structural characterisation and functional significance of transient protein-protein interactions. J. Mol. Biol. 325, 991–1018.
Nooren, I.M.A., and Thornton, J.M. (2003b). Diversity of proteinprotein interactions. EMBO J. 22, 3486–3492.
Papp, E., Nardai, G., Söti, C., and Csermely, P. (2003). Molecular chaperones, stress proteins and redox homeostasis. BioFactors 17, 249–257.
Park, S.G., Cha, M.K., Jeong, W., and Kim, I.H. (2000). Distinct physiological functions of thiol peroxidase isoenzymes in Saccharomyces cerevisiae. J. Biol. Chem. 275, 5723–5732.
Schröder, E., Littlechild, J.A., Lebedev, A.A., Errington, N., Vagin, A.A., and Isupov, M.N. (2000). Crystal structure of decameric 2-Cys peroxiredoxin from human erythrocytes at 1.7 Å resolution. Structure 8, 605–615.
Wood, Z.A., Schroder, E., Robin, H.J., and Poole, L.B. (2003). Structure, mechanism and regulation of peroxiredoxins. Trends Biochem. Sci. 28, 32–40.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
An, B.C., Lee, S.S., Lee, E.M. et al. New antioxidant with dual functions as a peroxidase and chaperone in Pseudomonas aeruginosa . Mol Cells 29, 145–151 (2010). https://doi.org/10.1007/s10059-010-0023-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10059-010-0023-1