• Biomedical Science Letters
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• v.15 no.1
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• pp.1-8
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• 2009

Human genomic DNA is continuously attacked by oxygen radicals originated from cellular metabolic processes and numerous environmental carcinogens. 2-deoxyribonolactone (dL) is a major type of oxidized abasic (AP) lesion implicated in DNA strand scission, mutagenesis, and formation of covalent DNA-protein crosslink (DPC) with DNA polymerase (Pol) ${\beta}$. We show here that human DNA polymerase (Pol)${\iota}$ and mitochondrial $Pol{\gamma}$ give rise to stable DNA-protein crosslink (DPC) formation that is specifically mediated by dL lesion. $Pol{\gamma}$ mediates DPC formation at the incised dL residue by its 5'-deoxyribose-5-phosphate (dRP) lyase activity, while $Pol{\gamma}$ cross links with dL thorough its intrinsic dRP lyase and AP lyase activities. Reactivity in forming dL-mediated DPC was significantly higher with $Pol{\gamma}$ than with $Pol{\iota}$. DPC formation by $Pol{\gamma}$, however, can be reduced by an accessory factor of $Pol{\gamma}$ holoenzyme that may attenuate deleterious effects of crosslink adducts on mitochondrial DNA. Comparative kinetic analysis of DPC formation showed that the rate of DPC formation with either $Pol{\iota}$ or $Pol{\gamma}$ was lower than that with $Pol{\beta}$. These results revealed that the activity of catalytic lyase in DNA polymerases determine the efficiency of DPC formation with dL damages. Irreversible crosslink formation of such DNA polymerases by dL lesions may result in a prolonged strand scission and a suicide of DNA repair proteins, both of which could pose a threat to the genetic and structural integrity of DNA.

• Molecules and Cells
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• v.39 no.1
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• pp.65-71
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• 2016

A challenge in the redox field is the elucidation of the molecular mechanisms, by which $H_2O_2$ mediates signal transduction in cells. This is relevant since redox pathways are disturbed in some pathologies. The transcription factor OxyR is the $H_2O_2$ sensor in bacteria, whereas Cys-based peroxidases are involved in the perception of this oxidant in eukaryotic cells. Three possible mechanisms may be involved in $H_2O_2$ signaling that are not mutually exclusive. In the simplest pathway, $H_2O_2$ signals through direct oxidation of the signaling protein, such as a phosphatase or a transcription factor. Although signaling proteins are frequently observed in the oxidized state in biological systems, in most cases their direct oxidation by $H_2O_2$ is too slow ($10^1M^{-1}s^{-1}$ range) to outcompete Cys-based peroxidases and glutathione. In some particular cellular compartments (such as vicinity of NADPH oxidases), it is possible that a signaling protein faces extremely high $H_2O_2$ concentrations, making the direct oxidation feasible. Alternatively, high $H_2O_2$ levels can hyperoxidize peroxiredoxins leading to local building up of $H_2O_2$ that then could oxidize a signaling protein (floodgate hypothesis). In a second model, $H_2O_2$ oxidizes Cys-based peroxidases that then through thiol-disulfide reshuffling would transmit the oxidized equivalents to the signaling protein. The third model of signaling is centered on the reducing substrate of Cys-based peroxidases that in most cases is thioredoxin. Is this model, peroxiredoxins would signal by modulating the thioredoxin redox status. More kinetic data is required to allow the identification of the complex network of thiol switches.

• Proceedings of the Korean Biophysical Society Conference
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• 2001.06a
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• pp.64-64
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• 2001

Escherichia coli has developed soxRS regulon to defend against toxicity of superoxide radical. SoxR, superoxide sensor, is oxidized by superoxide-generating agents or nitric oxide and oxidized SoxR activates the transcription of soxS gene. In order to find out the trans-acting factors regulating SoxR activity in vivo, soxS：：lacZ single copy operon fusion construct was prepared and random Tn10 insertional mutatons were performed.(omitted)

• Journal of Nutrition and Health
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• v.26 no.9
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• pp.1033-1048
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• 1993

This study was designed to describe the effect of the protein quality at different intake level of protein on the protein metabolism in the whole body of growing pigs with a simulation model. Varying to the protein level in feeds, four simulations were conducted. The feed protein level, represented as proportions of digestible protein to the metabolic energy (DP/ME, g/MJ), were 6-8, 11-13, 17-19, and 23-25 DP/ME, respectively. Two protein quality and six weeks of growth time were used at each simulation. The objective function for the simulations was protein deposition in the whole body, which was calculated from the experimental results. The parameters in the simulation were determined by the parameter estimation technique. The results obtained from the simulation were as follows: The protein synthesis and breakdown rates(g/day) in the whole body was increased with the increase of protein quality only at lower or required level of protein intake. They showed a parallel behavior in the course of growth, irrespective of quality and level of feed protein intake. The simulated protein deposition and protein synthesis showed a linear relationship between them at different protein quality and level. The affinity parameter showed a linear relationship between them at different protein quality and level. The affinity parameter showed that arginine, tryptophan and isoleucine were more efficient in the stimulation ofbody protein synthesis. Lysine and phenylalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, pheyalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, phenyalanine+tyrosine, and methionine+cystine were oxidized in larger magnitude than lysine and threonine. The oxidation parameter of most amino acids increased with the increase of protein intake beyond the requirement level, but not any more at highest protein intake level. Finally it was found that the improvement of feed protein quality at the lower or required level of protein intake increase protein deposition through a parallel increase of protein synthesis and breakdown.

• Journal of Microbiology and Biotechnology
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• v.31 no.12
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• pp.1732-1740
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• 2021

Tobacco etch virus protease (TEVp) is a useful tool for removing fusion tags, but wild-type TEVp is less stable under oxidized redox state. In this work, we introduced and combined C19S, C110S and C130S into TEVp variants containing T17S, L56V, N68D, I77V and S135G to improve protein solubility, and S219V to inhibit self-proteolysis. The solubility and cleavage activity of the constructed variants in Escherichia coli strains including BL21(DE3), BL21(DE3)pLys, Rossetta(DE3) and Origami(DE3) under the same induction conditions were analyzed and compared. The desirable soluble amounts, activity, and oxidative stability were identified to be reluctantly favored in the TEVp. Unlike C19S, C110S and C130S hardly impacted on decreasing protein solubility in the BL21(DE3), but they contributed to improved tolerance to the oxidative redox state in vivo and in vitro. After two fusion proteins were cleaved by purified TEVp protein containing double mutations under the oxidized redox state, the refolded disulfide-rich bovine enterokinase catalytic domain or maize peroxidase with enhanced yields were released from the regenerated amorphous cellulose via affinity absorption of the cellulose-binding module as the affinity tag.

• Journal of Photoscience
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• v.8 no.2
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• pp.43-53
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• 2001

a, a parameter that describes how effectively photoinactivated PS II units protect their functional neighbours; car, carotenoids; ΔpH, transthylakoid pH difference; D1 protein, psbA gene product in the PS II reaction centre; f, functional fraction of PS II: F$\_$v//F$\_$m/, the ratio of variable to maximum chlorophyll a fluorescence; k$\_$d/, rate coefficient for degradation of D1 protein; k$\_$i/ and k$\_$r/, rate coefficient for photoinactivation and repair of PS II, respectively; NADP+, oxidized nicontinamide adenine dinucleotide phosphate; P680, the primary electron donor in the PSII reaction centre; Ph, pheophytin; PS, photosystem; Q$\_$A/, first quinone acceptor of an electron in PS II; R$\_$s/, the gross rate of D1 protein synthesis.

• Journal of Life Science
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• v.5 no.2
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• pp.1-1
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• 1995

Casein or soybean protein was subjected to there action with caffeic acidtyrosinase system at 30-35$\circ$C, pH 6.8 with aeration for 5hr. The resulting brown proteins were washed with acetone until the washings were on longer colored. However, modified protein still retained a light brown. The effects of the modified proteins and brown compounds on male Wistar strain rats were studied by pair-feeding of a cholesterol-free diet for 14days. Significant decrease in protein digestibility for the rats fed with the modified proteins were observed. Weight gain and protein digestibility were not influenced by feeding brown compounds, but the feeding of brown compound from casein caused an enlargement of caecum. The concentrations of serum cholesterol and triglyceride in the rats fed with modified proteins and brown compounds were mostly unchanged against the rats fed with untreated proteins. These results suggest that the decrease in protein digestibility induced by enzymic browning-reaction did not cause the decrease in concentration of serum cholesterol.

• Journal of Life Science
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• v.13 no.5
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• pp.692-698
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• 2003

This study was designed to investigate the effects of ethyl acetate (EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle on membrane fluidity (MF), basal and induced oxygen radical (BOR and IOR), lipid peroxide (LPO) and oxidized protein (OP) as a oxidative stress, and lipofuscin (LF) in brain membranes of Sprague-Dawley (SD) rats. Male SD rats were fed basic diets (control) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45 days. MF was significantly increased (about 10%) in mitochondria of EtOAc-100 group. BOR and IOR formations in mitochondria were significantly inhibited (about 9∼10% and 17∼24%, respectively) in EtOAc-50 and EtOAc-100 groups, while BOR and IOR formations in microsomes were significantly inhibited (about 12∼17% and 12∼16%, respectively) in EtOAc-50 and EtOAc-100 groups compared with control group. LPO levels in mitochondria and microsomes were significantly inhibited (about 9∼l2% and 12∼19%, respectively) in EtOAc-50 and EtOAc-100 groups, whereas significant difference between OP or LF levels and control group in these membranes could not be obtained. These results suggest that administrations of ethyl acetate fraction of pine needle may play an effective role in an attenuating an oxidative stress and in increasing membrane fluidity.

• Journal of Life Science
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• v.13 no.5
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• pp.684-691
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• 2003

This study was designed to investigate the effects of ethyl acetate (EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle extract on membrane fluidity (MF), basal and induced oxygen radical (BOR and IOR), lipid peroxide(LPO) and oxidized protein (OP) as an oxidative stress, and lipofuscin(LF) in liver membranes of male Sprague-Dawley rats. Rats were fed basic diets (control) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45days. MFs were significantly increased (about 10%) in mitochondria of EtOAc-100 group compared with control group. BOR and IOR formations in mitochondria were significantly inhibited (about 12∼18% and 9 ∼l2%, respectively) in EtOAc-50 and EtOAc-100 groups, while BOR and IOR formations in microsomes were significantly inhibited (about 9∼l3% and 18∼19%, respectively) compared with control group. LPO levels were significantly inhibited (about 10% and 12∼13%, respectively) in mitochondria of EtOAc-100 and microsomes of EtOAc-50 and EtOAc-100 groups, whereas OP levels were significantly inhibited (about 13∼14%) in mitochondria of EtOAc-50 and EtOAc.-100 groups compared with control group. LF formations were significantly inhibited (about 10∼14%) in these three EtOAc groups. These results suggest that ethyl acetate fraction of pine needle may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity.

• BMB Reports
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• v.49 no.8
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• pp.443-448
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• 2016

The arginylation branch of the N-end rule pathway is a ubiquitin-mediated proteolytic system in which post-translational conjugation of Arg by ATE1-encoded Arg-tRNA-protein transferase to N-terminal Asp, Glu, or oxidized Cys residues generates essential degradation signals. Here, we characterized the ATE1^−/− mice and identified the essential role of N-terminal arginylation in neural tube development. ATE1-null mice showed severe intracerebral hemorrhages and cystic space near the neural tubes. Expression of ATE1 was prominent in the developing brain and spinal cord, and this pattern overlapped with the migration path of neural stem cells. The ATE1^−/− brain showed defective G-protein signaling. Finally, we observed reduced mitosis in ATE1^−/− neuroepithelium and a significantly higher nitric oxide concentration in the ATE1^−/− brain. Our results strongly suggest that the crucial role of ATE1 in neural tube development is directly related to proper turn-over of the RGS4 protein, which participate in the oxygen-sensing mechanism in the cells.