• Journal of the Korean Magnetic Resonance Society
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• v.8 no.1
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• pp.1-18
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• 2004

A thioredoxin from hyperthermophile, Methanococcus jannashii (MjTRX) was characterized by use of the differential scanning calorimetry to understand the mechanisms of thermodynamic stability. MjTRX has an unfolding transition temperature of 116.5$^{\circ}C$, although the maximum free energy of the unfolding (9.9 Kcal/mol) is similar to that of E. coli thioredoxin (ETRX, 9.0 Kcal/mol). However, the temperature of maximum stability is higher than ETRX by 20$^{\circ}C$, indicating that the unfolding transition temperature increased by shifting the temperature of maximum stability. MjTRX has lower enthalpy and entropy of the unfolding compared to ETRX maintaining a similar free energy of the unfolding. From the structure and the thermodynamic parameters of MjTRX, we showed that the unfolding transition temperature of MjTRX is increased due to the decreased entropy of the unfolding. Decreasing the unfolded state entropy and increasing the folded state entropy can decrease the entropy of the unfolding. In the case of MjTRX, the increased number of proline residues decreased the unfolded state entropy and the increased enthalpy in the folded state increased the folded state entropy.

• Biomolecules & Therapeutics
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• v.26 no.2
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• pp.121-129
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• 2018

Oxidized low-density lipoprotein (ox-LDL)-induced macrophage foam cell formation and apoptosis play critical roles in the pathogenesis of atherosclerosis. Thioredoxin-1 (Trx) is an antioxidant that potently protects various cells from oxidative stress-induced cell death. However, the protective effect of Trx on ox-LDL-induced macrophage foam cell formation and apoptosis has not been studied. This study aims to investigate the effect of recombinant human Trx (rhTrx) on ox-LDL-stimulated RAW264.7 macrophages and elucidate the possible mechanisms. RhTrx significantly inhibited ox-LDL-induced cholesterol accumulation and apoptosis in RAW264.7 macrophages. RhTrx also suppressed the ox-LDL-induced overproduction of lectin-like oxidized LDL receptor (LOX-1), Bax and activated caspase-3, but it increased the expression of Bcl-2. In addition, rhTrx markedly inhibited the ox-LDL-induced production of intracellular reactive oxygen species (ROS) and phosphorylation of p38 mitogen-activated protein kinases (MAPK). Furthermore, anisomycin (a p38 MAPK activator) abolished the protective effect of rhTrx on ox-LDL-stimulated RAW264.7 cells, and SB203580 (a p38 MAPK inhibitor) exerted a similar effect as rhTrx. Collectively, these findings indicate that rhTrx suppresses ox-LDL-stimulated foam cell formation and macrophage apoptosis by inhibiting ROS generation, p38 MAPK activation and LOX-1 expression. Therefore, we propose that rhTrx has therapeutic potential in the prevention and treatment of atherosclerosis.

• Biomolecules & Therapeutics
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• v.29 no.3
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• pp.321-330
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• 2021

Oxidative stress plays a crucial role in the development of neuronal disorders including brain ischemic injury. Thioredoxin 1 (Trx1), a 12 kDa oxidoreductase, has anti-oxidant and anti-apoptotic functions in various cells. It has been highly implicated in brain ischemic injury. However, the protective mechanism of Trx1 against hippocampal neuronal cell death is not identified yet. Using a cell permeable Tat-Trx1 protein, protective mechanism of Trx1 against hydrogen peroxide-induced cell death was examined using HT-22 cells and an ischemic animal model. Transduced Tat-Trx1 markedly inhibited intracellular ROS levels, DNA fragmentation, and cell death in H2O2-treatment HT-22 cells. Tat-Trx1 also significantly inhibited phosphorylation of ASK1 and MAPKs in signaling pathways of HT-22 cells. In addition, Tat-Trx1 regulated expression levels of Akt, NF-κB, and apoptosis related proteins. In an ischemia animal model, Tat-Trx1 markedly protected hippocampal neuronal cell death and reduced astrocytes and microglia activation. These findings indicate that transduced Tat-Trx1 might be a potential therapeutic agent for treating ischemic injury.

• Korean Journal of Applied Biomechanics
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• v.32 no.1
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• pp.31-36
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• 2022

Objective: The purpose of this study was to investigate muscle activation according to the four strap lengths in the TRX plank exercise to provide scientific and accurate data on effective training methods. Method: Twenty healthy men who had at least 6 months of weight training experience and could fully adjusted plank exercise, were participate in this study (age: 25.2 ± 3.7 yrs., height: 174.2 ± 3.9 cm., weight: 71.2 ± 9 kg). To pursue the study purpose, surface electrodes were attached to trunk muscles (pectoralis major, rectus abdominis, external oblique, internal oblique, erector spinae, latissimus dorsi) and lower extramity muscles (gluteus maximus, rectus femoris, gastrocnemius), and the muscle activity was measured using 11-channel electromyography equipment. In order to verify the muscle activation according to the four strap lengths during TRX plank exercise, an one-way ANOVA with repeated measure was used with statistical significance level set at as α=.05. Results: First, there were statistically significant differences in pectoralis major, rectus abdominis, external oblique, internal oblique, and erector spinae among TRX strap lengths. Second, there were statistically significant differences in gluteus maximus, rectus femoris, and gastrocnemius among TRX strap lengths. Third, even though no statistically significant difference found in latissimus dorsi, but increased muscle activation tendency was showed as the length of the strap increased. Conclusion: From the results of this study, it may be possible that TRX exercise prevent injuries and improve lower extremity muscle as well as trunk muscles by setting appropriate length of strap.

• BMB Reports
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• v.34 no.2
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• pp.139-143
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• 2001

Thioredoxin (Trx) is a redox protein possessing conserved sequence Cys-Gly-Pro-Cys in ail organisms. Trx acts as an electron donor of many proteins including thioredoxin peroxidase (TPx). Yeast Trx 2 has two redox active cysteine residues at positions 31 and 34. To investigate the redox activity of each cysteine, we generated mutants C31S, C34S, and C31S/C34S using site directed mutagenesis and examined the redox activity of Trx variants as an electron donor for yeast TPx enzymes. None of the three Cysmutated Trx proteins was active as a redox protein in the 5', 5'-dithiobis-(2-dinitrobenzoic acid) reduction under the condition of the presence of NADPH and thioredoxin reductase, and in the thioredoxin dependent peroxidase activity of yeast TPx II. C34S enhanced the glutamine synthetase protection activity of yeast TPx I, even though 100 times more protein was needed to exhibit the same activity to WT. The formation of a mixed disulfide intermediate between Trx and TPx II subunits was analyzed by SDS-PAGE. The mixed dieter form of TPx II was found only for C34S. These results suggest that Cys-31 more effectively acts as an electron donor for TPx enzymes.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.205-211
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• 2021

In order to test the functionality of Panax ginseng thioredoxin 1 (PgTRX1) isolated from fermented wild ginseng roots, a transient effect on physiological activity were performed over a short time frame using the Agrobacterium infiltration technique. The PgTRX1 gene isolated from fermented wild ginseng was confirmed to have a size of 579 bp, and the expression of PgTRX1 was the highest in the sample after 6 h of fermentation. As a result of constructing this gene and confirming the infiltration reaction mediated by Agrobacterium in tobacco leaves, it was found that the expression of the NbHSR203j gene was also induced as PgTRX1 expression increased. As a result of measuring the biological activity of the infiltration samples, the total phenol content increased by 35.45±1.84 to 49.01±1.84 ㎍ GAE/mL compared to the control, and the total flavonoid amount of 9.52±0.41 to 9.82±0.25 ㎍ QE/mL was slightly high. From these results, Agrobacterium-mediated PgTRX1 appears to be related to the hypersensitive response induction mechanism of plants and the production of secondary metabolites such as phenolic substances.

• Journal of Microbiology
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• v.44 no.1
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• pp.35-41
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• 2006

The unique gene encoding thioredoxin reductase (TrxR) was previously cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its expression was induced by oxidative stress. To elucidate tbe regulatory mechanism of the S. pombe TrxR gene, three fusion plasmids were generated using polymerase chain reaction: pYUTR20, pYUTR30, and pYUTR40. Plasmid pYUTR20 has an upstream region of 891 base pairs, pYUTR30 has 499 in this region, and pYUTR40 has an 186 bp upstream region. Negatively acting sequence is located between $-1,526\;\~\;-891bp$ upstream of the gene. The upstream sequence, responsible for the induction of TrxR by menadione (MD), is situated on the $-499\;\~\;-186bp$ region, which is also required for TrxR induction by mercuric chloride. The same region also appeared to be required for Pap1-mediated transcriptional regulation of the TrxR gene, which contains the two plausible Papl binding sites, TTACGAAT and TTACGCGA. Consistently, basal and inducible expression of the TrxR gene was markedly lower in the Pap1-negative TP108-3C cells than in wild-type yeast cells. In summary, up-regulation of the S. pombe TrxR gene is mediated by Pap1 via the transcriptional motif(s) located on the $-499\;\~\;-186bp$ region.

• Journal of The Korean Society of Integrative Medicine
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• v.12 no.2
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• pp.11-23
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• 2024

Purpose : The purpose of this study was to investigate the effect of exercise using a suspension device (sling and total body resistance exercise; TRX) on physical functional performance and postural balance. Methods : An experimental study comparing 2 different suspension exercise was conducted on 16 healthy college students. 16 subjects were assigned to two groups. They were classified into 8 sling group and 8 TRX group. Miniplus was used to evaluate physical functional performance. In this study, isokinetic resistance mode was used to compare and analyze seven movement patterns. Biorescue was used to evaluate postural balance. The intervention exercises in this study are as follows. Standing lean forward (SLF) using a sling and TRX was performed 3 times a week for 3 weeks. The SLF was held for 10 seconds and then rested for 15 seconds, repeated a total of 10 times for 3 sets. Results : In the TRX group, significant increases were observed in physical functional performance (p<.05). Among the differences between groups, significant differences were confirmed on the front of the right arm, the back of the left arm, and the back of the right arm. In the sling group, significant increases were observed in left, right, front, and overall dynamic balance (p<.05). A significant increase in posterior dynamic balance was confirmed in the TRX group (p<.05). There was no significant difference between groups. Conclusion : Based on the results of this study, TRX was effective in improving physical functional performance, while the sling was effective in enhancing postural balance. However, confirming the effectiveness of slings and TRX in the relationship between physical functional performance and postural balance proved inadequate. Therefore, additional research should be conducted to verify the effects of suspension.

• Biomolecules & Therapeutics
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• v.28 no.5
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• pp.443-455
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• 2020

The thioredoxin (Trx) system plays critical roles in regulating intracellular redox levels and defending organisms against oxidative stress. Recent studies indicated that Trx reductase (TrxR) was overexpressed in various types of human cancer cells indicating that the Trx-TrxR system may be a potential target for anti-cancer drug development. This study investigated the synergistic effect of auranofin, a TrxR-specific inhibitor, on sulforaphane-mediated apoptotic cell death using Hep3B cells. The results showed that sulforaphane significantly enhanced auranofin-induced apoptosis by inhibiting TrxR activity and cell proliferation compared to either single treatment. The synergistic effect of sulforaphane and auranofin on apoptosis was evidenced by an increased annexin-V-positive cells and Sub-G1 cells. The induction of apoptosis by the combined treatment caused the loss of mitochondrial membrane potential (ΔΨm) and upregulation of Bax. In addition, the proteolytic activities of caspases (-3, -8, and -9) and the degradation of poly (ADP-ribose) polymerase, a substrate protein of activated caspase-3, were also higher in the combined treatment. Moreover, combined treatment induced excessive generation of reactive oxygen species (ROS). However, treatment with N-acetyl-L-cysteine, a ROS scavenger, reduced combined treatment-induced ROS production and apoptosis. Thereby, these results deduce that ROS played a pivotal role in apoptosis induced by auranofin and sulforaphane. Furthermore, apoptosis induced by auranofin and sulforaphane was significantly increased through inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway. Taken together, the present study demonstrated that down-regulation of TrxR activity contributed to the synergistic effect of auranofin and sulforaphane on apoptosis through ROS production and inhibition of PI3K/Akt signaling pathway.

• Journal of Microbiology
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• v.44 no.5
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• pp.556-561
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• 2006

A differential display for the expression profiles of wild-type Cryphonectria parasitica and its virally-infected isogenic hypovirulent strain revealed several transcripts of interest, which evidenced significant matches with fungal genes of known function. Among which, we have further analyzed an amplified PCR product with significant sequence similarity to the known fungal stress-responsive thioredoxin gene from Neurospora crassa. The product of the cloned thioredoxin gene, CpTrx1, consists of 117 amino acids, with a predicted molecular mass of 13.0 kDa and a pI of 5.4. Sequence comparisons demonstrated that the deduced protein sequence of the CpTrx1 gene evidenced a high degree of homology to all known thioredoxins, with the highest degree of homology with trx1, a thioredoxin gene from Saccharomyces cerevisiae, and evidenced a preservation of the conserved hall markresidues (Trp-Cys-Gly-Pro-Cys) at the active site of thioredoxin. The E. coli-generated CpTRX1 manifested thioredoxin activity, according to the insulin reduction assay, which indicates that the cloned gene does indeed encode for the C. parasitica thioredoxin.