• Title/Summary/Keyword: Thiol-specific antioxidant protein

Search Result 17, Processing Time 0.028 seconds

Purification and Characterization of Thiol-Specific Antioxidant Protein from Human Liver: A Mer5-Like Human Isoenzyme

  • Cha, Mee-Kyung;Kim, Il-Han
    • BMB Reports
    • /
    • v.29 no.3
    • /
    • pp.236-240
    • /
    • 1996
  • A 23-kDa molecular mass of antioxidant protein was purified from human liver. This protein exhibited the preventive effect against the inactivation of glutamine synthetase by a metal-catalyzed oxidation system. This antioxidant activity was supported by a thiol-reducing equivalent such as dithiothreitol in a similar manner to that of the 25-kDa thiol-specific antioxidant protein (TSA) from human red blood cells (HR). However, a thioredoxin-linked peroxidase activity of thiol-specific antioxidant protein of human liver (HLTSA) (0.91 ${\mu}mol/min/nmol$ of HLTSA) was much lower than that of thiol-specific antioxidant protein of human red blood cells (HRTSA) (16.4 ${\mu}mol/min/nmol$ of HRTSA). This HLTSA is also immnologically distinct from HRTSA Amino acid sequences of the three tryptic peptides (P1, P2, P3) of HLTSA were found to be completely homologous to segments of the known Mer5-like protein, which belongs to the known TSA family.

  • PDF

Expression of Yeast Antioxidant Protein Gene in E. coil (대장균(JM 109)에서 효모 Thiol-Specific Antioxident 단백질의 발현)

  • Kim, Il-Han
    • The Journal of Natural Sciences
    • /
    • v.4
    • /
    • pp.1-10
    • /
    • 1991
  • The expression of the cloned Saccharomyces cerevisiae antioxidant protein gene in E. coil on plasmid was studied. Introduction of the DNA encoding yeast thiol-specific antioxidant protein into expression vector, PKK 223-3, having the tac promoter resulted in the production in E. coil of a Mr 27,000 polypeptide. This protein represented as much as about 1% of the bacterial soluble protein and showed the thiol-specific antioxidant properties. The bacterial protein showed physico-chemical properties indistringuishable from those of the yeast protein.

  • PDF

Purification and Characterization of TSA from Lumbricus terrestris (지렁이(Lumbricus terrestris)로부터 Thiol-Specific Antioxidant protein(TSA)의 분리 및 정제에 관한 연구)

  • Kwak, Byung-Koo;Kim, Il-Han;Cha, Mee-Kyung
    • The Journal of Natural Sciences
    • /
    • v.14 no.2
    • /
    • pp.55-65
    • /
    • 2004
  • A thiol-specific antioxidant(TSA) protein was purified from Earthworm, Lumbricus terrestris by DEAE-Cellulose, Phenl sepharose, Sephacryl S-200 gel filtration and HPLC S-300 Column Chromatography. This protein showed a thiol-specific antioxidant activity against inactivation of glutamine synthetase by a metal-catalysed oxidation system capable of generation reaction oxygen species. The molecular mass of the protein was determinated to be 51-kDa by SDS-polyacrylamide gel electrophores. Taken together, the purified TSA protein could be a new member of TSA family.

  • PDF

Purification and characterization of TPx from archeabacteria, Halococcus agglomeratus (고염 원시박테리아(Halococcus agglomeratus)에 존재하는 TPx 분리 및 생화학적 특성연구)

  • Choi, Yong-Soo;Cha, Mee-Kyung;Kim, Il-Han
    • The Journal of Natural Sciences
    • /
    • v.14 no.2
    • /
    • pp.67-82
    • /
    • 2004
  • A thiol-specific antioxidant protein (TSA or TPx) was purified from Halophilic archeabacteria Halococcus agglomeratus, by DEAE-Cellulose, Phnyl, sepharose, Sephadex G-75, Sephacryl S-100, Sephacryl S-200, and Q-Wepharose FF. This protein exhibited the preventeive effect against the inactivation of glutamine synthehase (GS) activity was support by a thiol-reducing equicalent such as dithiothreitol. TPx activity was maximal at NaCl concentration above 500mM. The molecular mass of the protein was determinated to be 22-kDa by SDS-PAGE. The TPx purified from Halococcus agglomeratus seems to be similar to other TPx family, except for the salt requirement for the maximal antioxidant activity.

  • PDF

Structural Mechanism for the Cellular Redox Regulation by the Thiol Specific Antioxidant Proteins

  • Park, Hee-Jeong;Kang, Sang-Won;Rhee, Sue-Goo;Ryu, Seong-Eon
    • Proceedings of the Korean Biophysical Society Conference
    • /
    • 1997.07a
    • /
    • pp.15-15
    • /
    • 1997
  • Recent studies indicate that hydrogen peroxide (H$_2$O$_2$), which is one of the reactive oxygen species involved in the oxidative stress, is an intracellular secondary messenger in the signal transduction. A novel family of thiol specific antioxidant (TSA) enzymes with a peroxidase activity shows no sequence homology to previously known antioxidant enzymes.(omitted)

  • PDF

Expression of Thiol-Dependent Protector Protein from Yeast Enhances the Resistance of Escherichia coli to Menadione

  • Park, Jeen-Woo;Ahn, Soo-Mi;Kim, Eun-Ju;Lee, Soo-Min
    • BMB Reports
    • /
    • v.29 no.6
    • /
    • pp.513-518
    • /
    • 1996
  • A soluble protein from Saccharomyces cerevisiae specifically provides protection against a thiolcontaining oxidation system but not against an oxidation system without thiol. This 25-kDa protein was thus named thiol-dependent protector protein (TPP). The role of TPP in the cellular defense against oxidative stress was investigated in Escherichia coli containing an expression vector with a yeast genomic DNA fragment that encodes TPP (strain YP) and a mutant in which the catalytically essential amino acid in the active site of TPP (Cys-47) has been replaced with alanine by site-directed mutagenesis (strain YPC47A). There was a distinct difference between these two strains in regard to viability, modulation of activities of superoxide dismutase and catalase, and the oxidative damage of DNA upon exposure to menadione. These results suggest that TPP may play a direct role in the cellular defense against oxidative stress by functioning as an antioxidant protein.

  • PDF

Characterization of Mouse Peroxiredoxin III Genomic DNA and Its Expression

  • Lee, Tae-Hoon;Kim, Sun-Uk;Lee, Kyung-Kwang;Yu, Dae-Yeul
    • Proceedings of the KSAR Conference
    • /
    • 2002.06a
    • /
    • pp.64-64
    • /
    • 2002
  • Peroxiredoxins (Prxs) possess protective activity against oxygen radicals generated by thiol-catalyzed oxidative systems. We already reported the genomic structure and its expression of mouse Prx Ⅰ, Ⅱ, and 1-Cys Prx. However, the Prx Ⅲ has not been determined. That was initially defined transiently expressed gene, mouse MER5, of murine erythroleukaemia cell differentiation. In addition, this protein was recently redefined a member of the thiol-specific antioxidant gene family. (omitted)

  • PDF

Identification of Proteins Affected by Iron in Saccharomyces cerevisiae Using Proteome Analysis

  • Lieu Hae-Youn;Song Hyung-Seok;Yang Seung-Nam;Kim Jae-Hwan;Kim Hyun-Joong;Park Young-Doo;Park Cheon-Seok;Kim Hae-Yeong
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.6
    • /
    • pp.946-951
    • /
    • 2006
  • To study the effect of iron on Saccharomyces cerevisiae, whole-cell proteins of Saccharomyces cerevisiae were extracted and subjected to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and differentially expressed proteins were identified. The proteins separated were further identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and were compared with a protein database. Of more than 300 spots separated by molecular weight and isoelectric points, 27 differentially expressed spots were identified. Ten proteins were found to be differentially expressed at high iron concentration. Triosephosphate isomerase (TPI), YDR533C hypothetical protein, superoxide dismutase (SOD), 60 kDa heat-shock protein (HSP60), pyruvate dehydrogenase beta subunit 1 (PDB1), and old yellow enzyme 2 (OYE2) were upregulated, whereas thiol-specific antioxidant (TSA), regulatory particle non-ATPase subunit 8 (RPN8), thiol-specific peroxiredoxin 1 (AHP1), and fructose-1, 6-bisphosphate adolase (FBA) were downregulated by iron. Based on the result, we propose that SOD upregulated by iron would protect the yeast from oxidative stress by iron, and that TSA downregulated by iron would render cells hypersensitive to oxidative stress.

Mechanism of Stress-dependent Structural Change of Yeast Prx (Yeast Prx의 스트레스의존 구조적 변화의 기작)

  • Kang, Ji-Seoun;Cheong, Gang-Won
    • Applied Microscopy
    • /
    • v.35 no.4
    • /
    • pp.16-23
    • /
    • 2005
  • Peroxiredoxins (Prxs) are a superfamily of thiol-specific antioxidant proteins present in all organism and involved in the hydroperoxide detoxification of the cell. To determine the structural organization of yeast-Prx, electron microscopic analysis was performed. The average images of yeast-Prxs revealed three different structure, i.e. spherical-shaped structure, ring-shaped structure and irregularly-shaped small particles. In order to analyze the conformational change of yeast-Prx by reduction and oxidation, Prxs were subjected to DTT and $H_2O_2$. In presence of DTT, yeast-Prx showed a high tendency to form a decamer. However, they changed into dimeric or spherical structure in the oxidized state. Here we also show ionic interaction between dimeric subunits is primarily responsible for yeast-Prx oligomerization.

Estrogen Regulate Neuroprotection and PDI Gene Expression in Ischemic Rat Brain

  • Yu, Seong-Jin;Kim, Do-Rim;Kim, Jee-Yun;Youm, Mi-Young;Lee, Chae-Kwan;Kang, Sung-Goo
    • Proceedings of the Korean Society of Developmental Biology Conference
    • /
    • 2003.10a
    • /
    • pp.69-69
    • /
    • 2003
  • Neuroprotective strategies have been appeared to be effective in a variety of stroke models. One of the major focuses has been related to the activities of estrogen. $17\beta$-estradiol valerate(EV) has been reported to exert neuroprotective effects when administered before an ischemic insult. The purpose of this study was to determine whether EV can protect against brain injury via estrogen receptor. Chronic and acute pretreatment can reduce the ischemic damage of focal cerebral ischemia in OVX rat, indicating that EV may be a new therapeutic class of drugs to prevent neuronal damage associated with cerebral ischemia. RNAs were extracted from the hippocampus of ovariectomized female rat with or without EV. Differential gene expression profiles were revealed(Bone morphogenetic protein type 1A receptor, Protein disulphide isomerase, cytochrome bc-1 complex core P, thiol-specific antioxidant protein). RT-PCR and in situ hybridization were used to validate the relative expression pattern obtained by the cDNA array. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) through the Biohealth Products Research Center(BPRC), Inje University, Korea

  • PDF