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Switching between the alternative structures and functions of a 2-Cys peroxiredoxin, by site-directed mutagenesis.
J Mol Biol. 2013 Nov 15; 425(22):4556-68.JM

Abstract

Members of the typical 2-Cys peroxiredoxin (Prx) subfamily represent an intriguing example of protein moonlighting behavior since this enzyme shifts function: indeed, upon chemical stimuli, such as oxidative stress, Prx undergoes a switch from peroxidase to molecular chaperone, associated to a change in quaternary structure from dimers/decamers to higher-molecular-weight (HMW) species. In order to detail the structural mechanism of this switch at molecular level, we have designed and expressed mutants of peroxiredoxin I from Schistosoma mansoni (SmPrxI) with constitutive HMW assembly and molecular chaperone activity. By a combination of X-ray crystallography, transmission electron microscopy and functional experiments, we defined the structural events responsible for the moonlighting behavior of 2-Cys Prx and we demonstrated that acidification is coupled to local structural variations localized at the active site and a change in oligomerization to HMW forms, similar to those induced by oxidative stress. Moreover, we suggest that the binding site of the unfolded polypeptide is at least in part contributed by the hydrophobic surface exposed by the unfolding of the active site. We also find an inverse correlation between the extent of ring stacking and molecular chaperone activity that is explained assuming that the binding occurs at the extremities of the nanotube, and the longer the nanotube is, the lesser the ratio binding sites/molecular mass is.

Authors+Show Affiliations

Department of Health, Life and Environmental Sciences, University of L'Aquila, Piazzale Salvatore Tommasi 1, 67100 L'Aquila, Italy. Electronic address: francesco.angelucci@univaq.it.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24021815

Citation

Angelucci, F, et al. "Switching Between the Alternative Structures and Functions of a 2-Cys Peroxiredoxin, By Site-directed Mutagenesis." Journal of Molecular Biology, vol. 425, no. 22, 2013, pp. 4556-68.
Angelucci F, Saccoccia F, Ardini M, et al. Switching between the alternative structures and functions of a 2-Cys peroxiredoxin, by site-directed mutagenesis. J Mol Biol. 2013;425(22):4556-68.
Angelucci, F., Saccoccia, F., Ardini, M., Boumis, G., Brunori, M., Di Leandro, L., Ippoliti, R., Miele, A. E., Natoli, G., Scotti, S., & Bellelli, A. (2013). Switching between the alternative structures and functions of a 2-Cys peroxiredoxin, by site-directed mutagenesis. Journal of Molecular Biology, 425(22), 4556-68. https://doi.org/10.1016/j.jmb.2013.09.002
Angelucci F, et al. Switching Between the Alternative Structures and Functions of a 2-Cys Peroxiredoxin, By Site-directed Mutagenesis. J Mol Biol. 2013 Nov 15;425(22):4556-68. PubMed PMID: 24021815.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Switching between the alternative structures and functions of a 2-Cys peroxiredoxin, by site-directed mutagenesis. AU - Angelucci,F, AU - Saccoccia,F, AU - Ardini,M, AU - Boumis,G, AU - Brunori,M, AU - Di Leandro,L, AU - Ippoliti,R, AU - Miele,A E, AU - Natoli,G, AU - Scotti,S, AU - Bellelli,A, Y1 - 2013/09/08/ PY - 2013/04/22/received PY - 2013/08/28/revised PY - 2013/09/02/accepted PY - 2013/9/12/entrez PY - 2013/9/12/pubmed PY - 2013/12/29/medline KW - 2-Cys peroxiredoxin KW - CS KW - HMW KW - LMW KW - PEG KW - Prx KW - SEC KW - TEM KW - citrate synthase KW - high molecular weight KW - host–parasite relationships KW - low molecular weight KW - molecular chaperone KW - moonlighting behavior KW - peroxiredoxin KW - polyethylene glycol KW - protein nanotube KW - size-exclusion chromatography KW - transmission electron microscopy KW - wild type KW - wt SP - 4556 EP - 68 JF - Journal of molecular biology JO - J Mol Biol VL - 425 IS - 22 N2 - Members of the typical 2-Cys peroxiredoxin (Prx) subfamily represent an intriguing example of protein moonlighting behavior since this enzyme shifts function: indeed, upon chemical stimuli, such as oxidative stress, Prx undergoes a switch from peroxidase to molecular chaperone, associated to a change in quaternary structure from dimers/decamers to higher-molecular-weight (HMW) species. In order to detail the structural mechanism of this switch at molecular level, we have designed and expressed mutants of peroxiredoxin I from Schistosoma mansoni (SmPrxI) with constitutive HMW assembly and molecular chaperone activity. By a combination of X-ray crystallography, transmission electron microscopy and functional experiments, we defined the structural events responsible for the moonlighting behavior of 2-Cys Prx and we demonstrated that acidification is coupled to local structural variations localized at the active site and a change in oligomerization to HMW forms, similar to those induced by oxidative stress. Moreover, we suggest that the binding site of the unfolded polypeptide is at least in part contributed by the hydrophobic surface exposed by the unfolding of the active site. We also find an inverse correlation between the extent of ring stacking and molecular chaperone activity that is explained assuming that the binding occurs at the extremities of the nanotube, and the longer the nanotube is, the lesser the ratio binding sites/molecular mass is. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/24021815/Switching_between_the_alternative_structures_and_functions_of_a_2_Cys_peroxiredoxin_by_site_directed_mutagenesis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(13)00559-7 DB - PRIME DP - Unbound Medicine ER -