• 제목/요약/키워드: peroxiredoxin 2

검색결과 66건 처리시간 0.031초

cDNA Sequence and mRNA Expression of a Novel Peroxiredoxin from the Firefly, pyrocoelia rufa

  • Jin, Byung-Rae;Lee, Kwang-Sik;Kim, Seong-Ryul;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제4권2호
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    • pp.101-107
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    • 2002
  • We describe here the cDNA sequence and mRNA expression of a novel family of the antioxidant protein, peroxiredoxin, from the firefly, Pyracoetia ruin. The 555 bp cDNA sequence codes for a 185 amino acid protein with a calculated molecular mass of approximately 21 kDa. The deduced protein of P. rufa peroxiredoxin gene contains two conserved cysteine residues. Alignment of the deduced protein of P. rufa peroxiredoxin gene showed 71.1% protein sequenceidentity to known insect Drosophila melanogaster peroxiredoxin. Northern blot analysis revealed that the P. rufa peroxiredoxin is specifically expressed in the fat body of P. rufa larvae.

Over-expressed Peroxiredoxin I Protects against Oxidative Damage in Mouse Embryonic Fibroblasts Lacking Peroxiredoxin II

  • Kim, Seong-Gon;Kim, Jae-Young;Ryoo, Zae-Young;Lee, Sang-Gyu
    • Biomolecules & Therapeutics
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    • 제19권4호
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    • pp.451-459
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    • 2011
  • Peroxiredoxins (Prxs) have a critical role in protecting cells against oxidative damage generated by reactive oxygen species (ROS). PrxI and PrxII are more than 90% homologous in their amino acid sequences, and both proteins reduce $H_2O_2$. In this study, an over-expression plasmid carrying PrxI was transfected into $PrxII^{-/-}$ mouse embryonic fibroblasts (MEFs) to investigate potential compensatory relationships between PrxI and PrxII. ROS levels induced by oxidative stress were increased in $PrxII^{-/-}$ MEFs as compared to wild-type MEFs. Moreover, exposure of $PrxII^{-/-}$ MEFs to $H_2O_2$ caused a reduction in cell viability of about 10%, and the proportion of cell death was increased compared to mock-treated $PrxII^{-/-}$ MEFs. However, transient over-expression of PrxI in $PrxII^{-/-}$ MEFs conferred increased resistance against the oxidative damage, as evidenced by increased cell viability and reduced intracellular ROS levels under $H_2O_2$ stress conditions. The findings suggest that over-expressed PrxI can partly compensate for the loss of PrxII function in PrxII-deficient MEFs.

애기장대 peroxiredoxins (Prxs)과 sulphiredoxin1 (Srx1)의 작용기작 (Working Mechanism of Peroxiredoxins (Prxs) and Sulphiredoxin1 (Srx1) in Arabidopsis thaliana)

  • 김민갑;수디 무하마드;박상렬;황덕주;배신철
    • 생명과학회지
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    • 제20권12호
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    • pp.1777-1783
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    • 2010
  • 식물체는 대사과정의 부산물로서 또는 생물학적으로 피해를 줄 수 있는 다양한 종류의 외부 스트레스에 직면했을 활성산소(Reactive Oxygen Species, ROS)를 생산한다. 이러한 oxidative 스트레스로부터 자신들을 보호하기 위하여 식물세포들은 다양한 종류의 항산화 단백질들을 보유하고 있다. 하지만 이들의 작용기작은 여전히 자세히 밝혀지지 않았다. Peroxiredoxins (Prxs)은 식물체에 광범위하게 존재하는 thiol-을 함유한 항산화 단백질로 N-말단에 존재하는 cysteine 잔기를 이용하여 hydrogen peroxide를 환원한다. 이러한 과정에서 peroxiredoxins의 활성부위인 cysteine 잔기는 선택적으로 cysteine sulfinic acid로 산화됨으로써 peroxidase activity의 불활성화를 일으킨다. 이러한 산화과정은 비가역적으로 일어난다. 최근 발견된 진핵생물들에 잘 보존된 sulphiredoxin (Srx1)이라 불리는 단백질은 cysteine-sulphinic acid를 환원시키는 기능을 지닌다. 본 논문에서는 애기장대에 존재하는 Prxs와 Srx의 기능에 대하여 서술할 예정이다.

Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • 제39권2호
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    • pp.133-141
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    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

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Survival of APC-mutant colorectal cancer cells requires interaction between tankyrase and a thiol peroxidase, peroxiredoxin II

  • Kang, Dong Hoon;Lee, Joanna H.S.;Kang, Sang Won
    • BMB Reports
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    • 제50권8호
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    • pp.391-392
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    • 2017
  • Overexpression of mammalian 2-Cys peroxiredoxin (Prx) enzymes is observed in most cancer tissues. Nevertheless, their specific roles in colorectal cancer (CRC) progression has yet to be fully elucidated. Here, a novel molecular mechanism by which PrxII/Tankyrase (TNKS) interaction mediates survival of adenomatous polyposis coli (APC)-mutant CRC cells was explored. In mice with an inactivating APC mutation, a model of spontaneous intestinal tumorigenesis, deletion of PrxII reduced intestinal adenomatous polyposis and thereby increased survival. In APC-mutant human CRC cells, PrxII depletion hindered PARP-dependent Axin1 degradation through TNKS inactivation. $H_2O_2-sensitive$ Cys residues in the zinc-binding domain of TNKS1 was found to be crucial for PARsylation activity. Mechanistically, direct binding of PrxII to ARC4/5 domains of TNKS conferred vital redox protection against oxidative inactivation. As a proof-of-concept experiment, a chemical compound targeting PrxII inhibited the growth of tumors xenografted with APC-mutation-positive CRC cells. Collectively, the results provide evidence revealing a novel redox mechanism for regulating TNKS activity such that physical interaction between PrxII and TNKS promoted survival of APC-mutant colorectal cancer cells by PrxII-dependent antioxidant shielding.

The Effects of Peroxiredoxin III on Human HeLa Cell Proliferation

  • Choi, Soonyoung;Kang, Sangwon
    • IMMUNE NETWORK
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    • 제3권4호
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    • pp.276-280
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    • 2003
  • Background: Peroxidases (Prx) of the peroxiredoxin family reduce hydrogen peroxide and alkyl hydroperoxides to water and alcohol respectively. Hydrogen peroxide is implicated as an intracellular messenger in various cellular responses such as proliferation and differentiation. And Prx I activity is regulated by Cdc-2 mediated phosphorylation. This work was undertaken to investigate the proliferation role of peroxiredoxin III as a member of Prx family in Prx III overexpressed HeLa cell line. Methods: To provide further evidence of proliferation, we selected Prx III stably expressed HeLa Tet-off cell lines. Cell proliferation was examined by using proliferation reagent WST-1 in the presence or absence of doxycycline. Prx III, 2-cys Prx enzymes exist as homodimer. The activation of Prx III heterodimer with induced and endogenous Prx III was examined by immunoprecipitation. Results: Immunoprecipitation analysis of the induced and endogenous Prx III with anti-myc showed that the induced wild type (WT) and dominant negative (DN) Prx III from HeLa Prx III Tet-off stable cell heterodimerized with endogenous Prx III each other. And the expression level of induced Prx III was examined after addition of doxycycline. By 72 hr, the expression level of induced Prx III was diminished gradually and the half-life of the induced wild type Prx III was approximately 17 hr. The proliferation experiment demonstrated that the relative proliferation value of induced and endogenous WT Prx III stable cell has no changes but the DN Prx III induced HeLa Tet-off stable cells were lower than endogenous Prx III. Conclusion: In conclusion, the HeLa dominant negative Prx III Tet-off stable cells were decreased the proliferation.

고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성 (Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato)

  • 박수영;류선화;권석윤;김종국;곽상수
    • Journal of Plant Biotechnology
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    • 제30권2호
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.

Characterization of Haemophilus influenzae Peroxiredoxins

  • Hwang, Young-Sun;Chae, Ho-Zoon;Kim, Kang-Hwa
    • BMB Reports
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    • 제33권6호
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    • pp.514-518
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    • 2000
  • Two open reading frames of Haemophilus influenzae, HI0572 and HI0751, showing homology to a yeast thioredoxin peroxidase II (TPx II) and an E. coli thiol peroxidase $P_{20}$, respectively, were cloned and expressed in E. coli, and then the proteins were subsequently purified and characterized. HI0751 protein showed the thioredoxin (Trx)-dependent peroxidase activity, whereas HI0572 protein showed glutathione-dependent peroxidase. The HI0572 is the first peroxiredoxin with glutathione peroxidase activity rather than thioredoxin peroxidase. Purified HI0572 and HI0751 proteins protected specifically the inactivation of glutamine synthetase by metal catalyzed oxidation (MCO) systems composed of $Fe^{3+}$, $O_2$ and mercaptans such as dithiothreitol, ${\beta}-mercaptoethanol$ and glutathione (GSH). Unlike the HI0751 protein, the HI0572 protein was more effective in protecting glutamine synthetase from inactivation by the $GSH/Fe^{3+}/O_2$ system. It seems that these unique properties of the HI0572 protein are due to the structure containing a glutaredoxin domain at it's C-terminal in addition to a peroxiredoxin domain.

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급성 부동 스트레스 후 척수 회색질에서 Peroxiredoxin I 및 III의 발현 변화 (Characterization of Peroxiredoxins in the Gray matter in the spinal cord after Acute Immobilization Stress)

  • 백남현;곽승수;이동석;이영호
    • Journal of Trauma and Injury
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    • 제19권2호
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    • pp.105-112
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    • 2006
  • Purpose: Many stresses produce reactive oxygen species and bring about mechanism of antioxidant reaction. Cytokine and a neurotransmitter through the cell membrane, as well as signal transduction through the cell membrane, are used for various pathological condition of the brain, such as neurodegenerative disease. There are several antioxidant enzymes in cells (superoxcide dismutase, glutathion peroxidasae, peroxiredoxin catalase, etc.) Methods: This study used single- or double-label immunohistochemical techniques to analyze mouse spinal neuron cells expressing Prx I and Prx III after acute mobilization stress. Results: Prx I was observed in dendritic cell of the gray matter of the spinal cord, and Prx III was observed in the cytoplasm of the GM of the spinal cord. Conclusion: The results of this study will help to explain differences of expression in the distributions of the peroxiredoxin enzymes of the spinal cord.

폐포상피세포, 대식세포를 비롯한 각종 세포주에서 H2O2에 의한 Peroxiredoxin 동위효소들의 산화에 따른 불활성화와 재생 (Oxidative Inactivation of Peroxiredoxin Isoforms by H2O2 in Pulmonary Epithelial, Macrophage, and other Cell Lines with their Subsequent Regeneration)

  • 오윤정;김영선;최영인;신승수;박주헌;최영화;박광주;박래웅;황성철
    • Tuberculosis and Respiratory Diseases
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    • 제58권1호
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    • pp.31-42
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    • 2005
  • 배 경 : peroxiredoxins는 거의 모든 생명체에 공통적으로 보존되어 있으며, 최근에 발견된, 특이한 peroxidases로 인체에서 6가지 동위효소가 알려져 있으며, 산화스트레스에 대한 방어역할을 담당하고, $H_2O_2$신호전달 과정에서 중요한 조절 역할을 한다. peroxiredoxin은 $H_2O_2$ 처리 과정 중에서 자신이 산화되어 불활성화 되는데, 산화된 후 다시 재생되는 것으로 보고되나 그 생리적은 의미는 분명하지 않다. 이에 저자들을 폐상 피세포주, 대식세포주, 폐포모세혈관 내피세포주 및 기타 섬유모세포주 들에서 $H_2O_2$ 에 의한 Prx의 산화과정과 재생을 알아보고자 하였다. 방 법 : 수술 환자에서 적출한 정상 폐조직과, 세포주로는 평상시 산화 스트레스에 노출이 많을 것으로 예상되는 세포들로써, 폐포상피세포의 I 형 및 II 형 세포에서 기원한 A549, WI 26, Raw 264.7, Rat2,및 폐포 모세혈관 내피세포주 등을 이용하여 이를 $50{\mu}M$. $100{\mu}M$, $500{\mu}M$$H_2O_2$로 산화시켜 불활성화 한 후, 추적관찰 하였으며, 시간대 별로(0. 10, 30, 60, 120, 240, 480 분) 수확하여, 이를 1차원 non-reducing SDS-PAGE 및 2차원 전기영동로 분리 후, silver stain 과 Western blot으로 분석 하였다. 결 과 : 1. 실험에 사용된 모든 세포주에서, $H_2O_2$ 농도에 비례하여 peroxiredoxin I, II, III 의 불활성화를 관찰할 수 있었고, 10분에 최고로 불활성화되었다. 2. 산화된 이후, 30분경부터 peroxiredoxin 의 재생이 관찰되기 시작 하였으며, 2시간 이후부터 확연하였다. 3. 다시 재생된 peroxiredoxin은 $H_2O_2$투여로서, 다시 불활성화되어, 재생된 Prx 가 활성을 지닌 단백질임을 알 수 있었다. 4. 재생의 속도는 사용된 세포주마다 차이가 있었으며 (A549 >Raw 264.7 >$Rat_2$ >WI26), 단백질 합성억제제인 cycloheximide ($10{\mu}g/ml$) 존재 하에서도 변함 없이 관찰되었다. 결 론 : 세포 내에는 산화되어 불활성화된 peroxiredoxin을 재생하는 체계가 존재 하며, 이는 활성부위 cysteine을 갖는 다른 단백질에도 공통적으로 적용될 수 있는 분자 스위치일 가능성이 높으며, 산화에 의한 신호전달과정이나, 질병 모델에서 Prx 단백의 재생 체계의 이상과 병인에 관한 추가적인 연구가 필요할 것으로 사료된다.