• Title/Summary/Keyword: 유도단백질

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Characterization of Immunogens of Infectious Hematopoietic Necrosis Virus Isolated in Korea (전염성 조혈기 괴사 바이러스(IHNV)의 항원 유도 단백질 특성)

  • Park, Myoung-Ae;Sohn, Sang-Gyu;Park, Jeong-Woo;Jeong, Young-Kee
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.13-22
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    • 1994
  • To identify the immunogens of a PRT strain of Infectious Hematopoietci Necrosis Virus (IHNV) isolated from cultrued fish in Korea (Park et al, 1993). a panel of 4 monoclonal antibodies (MAbs) against IHNV-PRT strain and two polyclonal antisera from rainbow trout survived IHN disease were prepared. Proteins of purified IHNV-PRT strain were analysed on 10% SDS-PAGE and transferred onto NC paper and were incubated with the antibody solutions. With the polyclonal antibodies, four bands ($M_1$, $M_2$, G and 90Kd) were detected and the band density was in the order of $M_2$ > 90Kd > $M_1$ > G. However, with the MAbs, only two bands(G and 90Kd) were detected. The origin of 90Kd protein was not clear but maybe cell. All the results represented that among the five proteins of IHNV-PRT strain (Park et al., 1993), $M_2$, $M_1$ and G proteins were immunogens and $M_2$ protein was the strongest one.

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Proteomics of Protein Expression Profiling in Tissues with Different Radiosensitivity (Proteomics를 이용한 마우스 조직에서의 방사선 감수성 조절 단백질의 탐색)

  • An, Jeung-Hee;Kim, Ji-Young;Seong, Jin-Sil
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.298-306
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    • 2004
  • Purpose: The purpose of this study was to identify Radiosensitivity of proteins in tissues with different radiosensitivity. Materials and Methods: C3H/HeJ mice were exposed to 10 Gy. The mice were sacrifiud 8 hrs after radiation. Their spleen and liver tissues were collected and analyzed histologicaly for apoptosis. The expressions of radiosusceptibillty protein were analyzed by 2-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Resilts: The Peak of apoptosis levels were $35.3{\pm}1.7{\%}$ in spleen and $0.6{\pm}0.2{\%}$ in liver at 8 hrs after radiation. Liver, radioresistant tissues, showed that the levels of ROS metabolism related to proteins such as cytochromm c, glutathione S transferase, NADH dehydrogenase, riken cDNA and peroxiredoxin Vl increased after radiation. The expression of cytochrome c increased significantly in spleen and liver tissues after radiation. In spleen, radiosensitivity tissue, the identified proteins showed a significantly quantitative alteration following radiation. It was categorized as signal transduction, apoptosis, cytokine, Ca signal related protein, stress-related protein, cytoskeletal regulation, ROS metabolism, and others. Conclusion: Differences of radiation-induced apoptosis by tissues specifted were coupled with the induction of related radiosensitivity and radioresistant proteins. The result suggests that apoptosis relate protein and redox proteins play important roles in this radiosusceptibility.

The Effect of Growth Regulators and Light Quality on the Changes in Protein Pattern of Callus from Intergeneric Protoplast Fusion between Nicotiana tabacum and Solanum nigrum (Nicotiana tabacum과 Solanum nigrum의 속간 원형질체 융합에서 유도된 캘러스의 단백질 양태변화에 미치는 생장조절제 및 광선의 효과)

  • 김영상;이동희
    • Journal of Environmental Science International
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    • v.3 no.2
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    • pp.141-155
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    • 1994
  • The effect of growth regulators (NAA, BA and $ extrm{GA}_3$) and light (blue, red and far-red) on the changes in total protein and thylakoid membrane protein pattern of callus from intergeneric protoplast fusion between Nicotiana tabacum and Solanum nigrw were investigated. When the callus were irradiated with different wavelengths of light, blue and red light accelerated the synthesis of total proteins and thylakoid membrane proteins. Particularly, red light led to an increase in the protein synthesis compared to blue light. When the callus were subjected to various combinations of growth regulators, NAA+$ extrm{GA}_3$ and NAA+BA treatments induced remarkable increase of total proteins and thylakoid membrane proteins accumulation, particularly in the combination of NAA+$ extrm{GA}_3$. NAA.$ extrm{GA}_3$ treatment with irradiation of red ligh showed highest value in the accumulation of total proteins and thylakoid membrane proteins. We conclude that simultaneous application of red light and NAA+$ extrm{GA}_3$ treatment may induce synergistic effect in the synthesis of total proteins and thylakoid membrane Proteins.

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Studies on the Transformation of Crop Plants. IV. Biochemical Characteristics of Embryogenic Callus in Rice (곡물류의 형질전환 유도에 관한 연구 (IV) 벼 배발생 세포의 생화학적 특징)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.377-382
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    • 1993
  • Rice (Oryza saliva L.) calli containing both embryogenic callus (EC) and non embryogenic callus (NEC) regions were initiated from the mature seed on MS medium supplemented with 2.0 mg/L 2,4-D, 0.5 mg/L kinetin. The calli were developed into two callus type which can be distinguished by visual examination depending on color and appearance. In order to illucidate the polypeptide composition between EC and NEC, the total protein extracted from two types of callus was analysed by electrophoresis. By one-dimesional anlaysis of SDS-PAGE and Isoelectric focusing, several protein bands showed quantitative and qualitative difference in each type of callus. The further analysis of the total protein with two-dimensional electrophoresis showed at least 20 EC specific protein and 10 NE specific protein. Also 3 specific protein spots showing micro heterogeneity of 90, 65, 50 kD were detected in EC, while a series of acidic heterologous protein spots were visualized in NEC.in NEC.

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The Purification and Immunogenicity of TB-14 Recombinant Protein of Mycobacterium tuberculosis (결핵균 특이 TB-14 재조합 단백질의 분리 및 세포성 면역반응에 미치는 영향)

  • Song, Ho-Yeon;Kim, Young-Hee;Kim, Chang-Hwan;Min, Young-Ki;Kim, Dae-Joong;Ko, Kwang-Kjune
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.3
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    • pp.239-247
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    • 2006
  • Background: Culture filtrate proteins secreted by mycobacteria are thought to play an important role in inducing protective immunity and to develop new methods for diagnosing tuberculosis. Methods: A culture filtrate protein of M. avium that was strongly reactive with goat antiserum against M. intracellulare was constructed. Its homologous protein (TB-14) in M. tuberculosis was cloned, expressed and purified. The inductions of IFN-${\gamma}$ stimulated with $10{\mu}g$ of TB-14 recombinant protein and $10{\mu}g$ PPD were estimated by using whole bloods from seven PPD (-) subjects, seven PPD (+) healthy volunteers and nine tuberculosis patients. Results: M. avium culture filtrate protein was confirmed as a hypothetical protein that was termed contig 116. A novel 14-kDa recombinant protein (TB-14) of M. tuberculosis was composed of 148 amino acids, including 30 amino acids of the signal peptide, and it showed 78% homology with M. avium. In the PPD (+) healthy volunteers, recombinant TB-14 protein strongly induced the secretion of IFN-${\gamma}$ in whole blood cultures. Conclusion: These results suggest that TB-14 recombinant protein might play an important role in inducing cell-mediated immunity against tuberculosis. Furthermore, TB-14 protein antigen and its antiserum will be available for the development of new diagnostic tools for tuberculosis.

$\beta$-catenin에 의한 신호전달과 그 역할 ($\beta$-catenin은 세포의 감초인가\ulcorner)

  • 정선주
    • The Zoological Society Korea : Newsletter
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    • v.18 no.1
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    • pp.16-25
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    • 2001
  • Wnt signaling의 주요 분자인 $\beta$-catenin의 기능과 조절에 관한 연구, 특히 TCF family 단백질과 함께 작용하는 신호전달에 관한 연구가 최근에 활발히 진행되고 있다. $\beta$-catenin 단백질은 Drosophila나 Xenopus의 발생초기에 중요한 역할을 한다는 것이 알려져 있고 Wnt (Wingless) 단백질에 의하여 활성화되는 신호전달 과정에 관여한다고 알려져 있으므로, TCF 단백질들이 Wnt signalling pathway에 작용한다는 것을 의미한다. 즉, $\beta$-catenin/TCF complex는 발생초기의 세포의 운명을 결정하는 세포의 분화에 중요하리라 생각된다. 또한 $\beta$-catenin/TCF complex는 세포의 암화에도 중요하다는 것이 보고되었다. 정상세포의 경우, $\beta$-catenin은 APC 라는 tumor suppressor에 의하여 결합하고 단백질의 분해가 유도되어 핵 안의 TCF와 결합하지 못하는데, 암세포의 경우 APC가 결실되었거나 $\beta$-catenin의 양이 과도하게 발현되어 암세포화 되는 것으로 보인다. 즉, $\beta$-catenin은 일종의 oncogene으로 작용하는 단백질이며, 그 작용에 필수적인 전사인자가 TCF라는 것이다. 특히, 대장암세포에서 이 $\beta$-catenin/TCF complex에 의해 활성화되는 유전자로서 c-myc과 cyclin Dl 등이 있는 것으로 보아, $\beta$-catenin/TCF 단백질은 세포의 증식 및 사멸에 관여하는 단백질들의 발현을 조절하는 매우 중요한 인자라고 생각된다.

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Induction of Apoptosis in HT-29 Human Colon Cancer Cells by the Pepper Component Piperine (후추의 주요 성분인 Piperine의 대장암세포 세포사멸 유도 효과)

  • Kim, Eun-Ji;Park, Hee-Sook;Shin, Min-Jeong;Shin, Hyun-Kyung;YoonPark, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.442-450
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    • 2009
  • Piperine is an alkaloid-amine found in pepper and has been reported to have anticarcinogenic properties. To explore the possibility that piperine has cancer chemopreventive and chemotherapeutic effects in colon cancer, we examined whether piperine inhibits the growth of HT-29 human colon cancer cells and investigated the mechanisms for this effect. Cells were cultured with various concentrations ($0{\sim}40{\mu}M$) of piperine. Piperine decreased the cell viability and induced apoptosis of HT-29 cells. Western blot analysis of total cell lysates revealed that piperine decreases the protein levels of Bcl-2, Mcl-1, and intact Bid but increases Bik levels. Piperine increased the percentage of cells with depolarized mitochondrial membrane, and the release of cytochrome c into cytoplasm. Piperine induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3 and increased the Fas levels. In addition, piperine significantly decreased the protein levels of survivin. The present results indicate that piperine inhibits the growth of HT-29 colon cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl-2 family proteins, increase the activation of caspases, and decrease survivin levels. Overall, our findings suggest that piperine has cancer chemotherapeutic effects in colon cancer.

Induction of Apoptosis in FRTL-5 Thyroid Cells by Okadaic Acid (Okadaic Acid에 의한 FRTL-5 갑상선 세포주의 Apoptosis 유도)

  • Cho Ji-Hyoung;Chung Ki-Yong;Park Jong-Wook
    • Korean Journal of Head & Neck Oncology
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    • v.18 no.2
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    • pp.142-149
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    • 2002
  • Objectve : Okadaic acid is a specific inhibitor of serine/threonine protein phosphatase 1 and 2A. In order to know the mechanism of apoptosis induced by okadaic acid, we treated FRTL-5 thyroid cells with okadaic acid and measured the changes of important proteins that are involved in apoptosis. Materials and Methods: We measured caspase 3 activity, $PLC-{\gamma}1$ degradation, the expression of XIAP, cIAP1, cIAP2, and cytochrome c release in okadaic acid-treated FRTL-5 thyroid cells. Results: Okadaic acid-induced caspase 3 activation and $PLC-{\gamma}1$ degradation and apoptosis were dose-dependent with a maximal effect at a concentration of 80 nmol and time-dependent with a maximal effect at 24 hours after treatment. The elevated caspase 3 activity in okadaic acid treated FRTL-5 thyroid cells are correlated with down-regulation of XIAP and cIAP1, but not cIAP2. General and potent inhibitor of caspases, z-VAD-fmk. abolished okadaic acid-induced caspase 3 activity and $PLC-{\gamma}1$ degradation. The release of cytochrome c in okadaic acid-induced FRTL-5 thyroid cells was dose-dependent with a maximal effect at a concentration of 80 nmol. Conclusions: These findings suggest that mechanism of okadaic acid-induced apoptosis is associated with cytochrome c release and increase of caspase 3 activation in FRTL-5 thyroid cells.

Mcl-1 is a Binding Partner of hNoxa (Mcl-1 단백질은 Noxa 단백질의 결합 파트너이다.)

  • Park, Sun-Young;Kim, Tae-Hyoung
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1063-1067
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    • 2007
  • The Bcl-2 family proteins play critical roles in regulation of apoptosis, and the balanced interaction of pro- and anti-death members is a key factor in determining the cell fate. Noxa, a BH3-only Bcl-2-family member, has been originally identified as a target gene of p53. To understand the mechanism by which human Noxa (hNoxa) regulates the cell death, we screened the hNoxa binding partner using the yeast two hybrid screening and found that anti-death protein Mcl-1 binds to hNoxa. The binding of hNoxa to Mcl-1 was confirmed by immunoprecipitation in human colon cancer cell line HCT 116 cells. Mcl-1 significantly inhibited the hNoxa-induced cell death in HCT 116 cells. During the cell death induced by hNoxa, Mcl-1 protein was degraded. Its degradation was inhibited by z-VAD-fmk, a pancaspase inhibitor, suggesting caspase is responsible for Mcl-1 degradation in response to hNoxa. Together, the results indicate that hNoxa binds to Mcl-1 that is degraded by cas-pases during hNoxa-induced cell death.