Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance

An, Byung Chull; Lee, Seung Sik; Lee, Jae Taek; Hong, Sung Hwan; Wi, Seung Gon; Chung, Byung Yeoup

doi:10.1007/s10059-011-1047-x

Cited by 9 publications

(7 citation statements)

References 27 publications

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“…However, evolution has favoured a multifunctional 2-CysPrx protein that switches between the high-affinity but low-turnover peroxidase function and the low-efficiency chaperone function. There is a need in plants, but also in mammals and other organisms, to explore the reasons for this delicate multifunctionality beyond the reported positive effects of, for example, overexpression of 2-CysPrx on oxidative stress and heat tolerance (An et al , 2011; Kim et al , 2011). …”

Section: Discussionmentioning

confidence: 99%

The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone

König

Galliardt

Jütte

et al. 2013

Journal of Experimental Botany

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2-Cysteine peroxiredoxins (2-CysPrxs) are ubiquitous and highly abundant proteins that serve multiple functions as peroxidases, chaperones, and thiol oxidases and in redox-dependent cell signalling. The chloroplast protein plays a role in seedling development and protection of the photosynthetic apparatus. This study aimed to unequivocally link conformation and function. To this end, a set of non-tagged site-directed mutagenized At2-CysPrx variants was engineered, which mimicked the conformational states and their specific functions: hyperoxidized form (C54D), reduced form (C54S, C176S), oxidized form (C54DC176K), phosphorylated form (T92D), reduced ability for oligomerization by interfering with the dimer–dimer interface (F84R) and a C-terminally truncated form [ΔC (–20 aa)]. These variants were fully or partly fixed in their quaternary structure and function, respectively, and were analysed for their conformational state and peroxidase and chaperone activity, as well as for their sensitivity to hyperoxidation. The presence of a His6-tag strongly influenced the properties of the protein. The ΔC variant became insensitive to hyperoxidation, while T92D and F84R became more sensitive. The C54D variant revealed the highest chaperone activity. The highest peroxidase activity was observed for the F84R and ΔC variants. Efficient interaction with NADP-dependent thioredoxin reductase C depended on the presence of Cys residues and the C-terminal tail. The results suggest that the structural flexibility is important for the switch between peroxidase and chaperone function and that evolution has conserved the functional switch instead of maximizing a single function. These variants are ideal tools for future conformation-specific studies in vivo and in vitro.

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Section: Discussionmentioning

confidence: 99%

The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone

König

Galliardt

Jütte

et al. 2013

Journal of Experimental Botany

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“…In our previous studies, radiation-mediated changes in the structure and function of the protein was recorded by determining the changes in the molecular properties of 2-cysteine peroxiredoxins (2-Cys Prxs), which have dual enzymatic functions of a peroxidase and a molecular chaperone [ 11 , 12 , 13 , 14 ]. Exposure of 2-Cys Prxs to ionizing radiations, such as, γ rays and electron beams, greatly increased the holdase chaperone activity increased by 3–4-fold compared to non-irradiated protein, but peroxidase activity declined with increasing radiation doses [ 12 , 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

Enhancement of Chaperone Activity of Plant-Specific Thioredoxin through γ-Ray Mediated Conformational Change

Lee

Jung

Park

et al. 2015

IJMS

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AtTDX, a thioredoxin-like plant-specific protein present in Arabidospis is a thermo-stable and multi-functional enzyme. This enzyme is known to act as a thioredoxin and as a molecular chaperone depending upon its oligomeric status. The present study examines the effects of γ-irradiation on the structural and functional changes of AtTDX. Holdase chaperone activity of AtTDX was increased and reached a maximum at 10 kGy of γ-irradiation and declined subsequently in a dose-dependent manner, together with no effect on foldase chaperone activity. However, thioredoxin activity decreased gradually with increasing irradiation. Electrophoresis and size exclusion chromatography analysis showed that AtTDX had a tendency to form high molecular weight (HMW) complexes after γ-irradiation and γ-ray-induced HMW complexes were tightly associated with a holdase chaperone activity. The hydrophobicity of AtTDX increased with an increase in irradiation dose till 20 kGy and thereafter decreased further. Analysis of the secondary structures of AtTDX using far UV-circular dichroism spectra revealed that the irradiation remarkably increased the exposure of β-sheets and random coils with a dramatic decrease in α-helices and turn elements in a dose-dependent manner. The data of the present study suggest that γ-irradiation may be a useful tool for increasing holdase chaperone activity without adversely affecting foldase chaperone activity of thioredoxin-like proteins.

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“…The PCR products of the single and double mutations were collected and purified, treated with DpnI (Enzynomics, Korea) to remove the original DNA, and then transformed into DH5α cells. The single and double mutations were confirmed by nucleotide sequencing after plasmid isolation ( An et al, 2011 ; 2015 ).…”

Section: Methodsmentioning

confidence: 99%

Epigenetic and Glucocorticoid Receptor-Mediated Regulation of Glutathione Peroxidase 3 in Lung Cancer Cells

An¹,

Jung²,

Park³

et al. 2016

Molecules and Cells

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Glutathione peroxidase 3 (GPx3), an antioxidant enzyme, acts as a modulator of redox signaling, has immunomodulatory function, and catalyzes the detoxification of reactive oxygen species (ROS). GPx3 has been identified as a tumor suppressor in many cancers. Although hyper-methylation of the GPx3 promoter has been shown to down-regulate its expression, other mechanisms by which GPx3 expression is regulated have not been reported. The aim of this study was to further elucidate the mechanisms of GPx3 regulation. GPx3 gene analysis predicted the presence of ten glucocorticoid response elements (GREs) on the GPx3 gene. This result prompted us to investigate whether GPx3 expression is regulated by the glucocorticoid receptor (GR), which is implicated in tumor response to chemotherapy. The corticosteroid dexamethasone (Dex) was used to examine the possible relationship between GR and GPx3 expression. Dex significantly induced GPx3 expression in H1299, H1650, and H1975 cell lines, which exhibit low levels of GPx3 expression under normal conditions. The results of EMSA and ChIP-PCR suggest that GR binds directly to GRE 6 and 7, both of which are located near the GPx3 promoter. Assessment of GPx3 transcription efficiency using a luciferase reporter system showed that blocking formation of the GR-GRE complexes reduced luciferase activity by 7–8-fold. Suppression of GR expression by siRNA transfection also induced down-regulation of GPx3. These data indicate that GPx3 expression can be regulated independently via epigenetic or GR-mediated mechanisms in lung cancer cells, and suggest that GPx3 could potentiate glucocorticoid (GC)-mediated anti-inflammatory signaling in lung cancer cells.

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Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance

Cited by 9 publications

References 27 publications

The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone

The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone

Enhancement of Chaperone Activity of Plant-Specific Thioredoxin through γ-Ray Mediated Conformational Change

Epigenetic and Glucocorticoid Receptor-Mediated Regulation of Glutathione Peroxidase 3 in Lung Cancer Cells

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