• 제목/요약/키워드: p56

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Transcription, Translation, and Immunolocalization of ODVP-6E/ODV-E56 and p74 Proteins: Two Highly Conserved ODV-associated Envelope Proteins of Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
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    • 제38권1호
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    • pp.65-70
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    • 2005
  • Choristoneura fumiferana granulovirus (ChfuGV) infection results two types of enveloped virions: Occlusion-derived virus (ODV) and budded virus (BV). Structural proteins ODVP-6E/ODV-E56 and p74 are two major conserved ODV-associated proteins that may be involved in the initiation of viral infection cycle in susceptible host insect larvae. This study presents the characterization of ChfuGV odvp-6e/odv-e56 and p74 transcription and translation as well as immunolocalization of these proteins in the occluded ChfuGV virion. Our results revealed that the transcription of odvp-6e/odv-e56 and p74 genes, both, start at 24 hours post infection (h p.i.). Using monospecific polyclonal antibodies made against ODVP-6E/ODV-E56 and p74 we demonstrated that these proteins are both expressed late in infection (24 h p.i.). Immunogold labeling using antisera against ODVP-6E/ODV-E56 and p74 proteins demonstrated that ODVP-6E/ODV-E56 and p74 proteins are both associated with the ODV envelop of ChfuGV.

Tim-3 Expression by Peripheral Natural Killer Cells and Natural Killer T Cells Increases in Patients with Lung Cancer - Reduction after Surgical Resection

  • Xu, Li-Yun;Chen, Dong-Dong;He, Jian-Ying;Lu, Chang-Chang;Liu, Xiao-Guang;Le, Han-Bo;Wang, Chao-Ye;Zhang, Yong-Kui
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권22호
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    • pp.9945-9948
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    • 2014
  • Background: The purpose of this study was to investigate Tim-3 expression on peripheral CD3-CD56+ natural killer (NK) cells and CD3+CD56+ natural killer T (NKT) cells in lung cancer patients. Materials and Methods: We analyzed Tim-3+CD3-CD56+ cells, Tim-3+CD3-$CD56^{dim}$ cells, Tim-3+CD3-$CD56^{bright}$ cells, and Tim-3+CD3+CD56+ cells in fresh peripheral blood from 79 lung cancer cases preoperatively and 53 healthy controls by flow cytometry. Postoperative blood samples were also analyzed from 21 members of the lung cancer patient cohort. Results: It was showed that expression of Tim-3 was significantly increased on CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in lung cancer patients as compared to healthy controls (p=0.03, p=0.03 and p=0.04, respectively). When analyzing Tim-3 expression with cancer progression, results revealed more elevated Tim-3 expression in CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in cases with advanced stages (III/IV) than those with stage I and II (p=0.02, p=0.04 and p=0.01, respectively). In addition, Tim-3 expression was significantly reduced on after surgical resection of the primary tumor (p<0.01). Conclusions: Tim-3 expression in natural killer cells from fresh peripheral blood may provide a useful indicator of disease progression of lung cancer. Furthermore, it was indicated that Tim-3 might be as a therapeutic target.

Jurkat T 세포에서 Ginsenoside-Rg1이 p561ck Kinase 활성과 세포증식에 미치는 영향 (Stimulatory Effects of Ginsenoside-Rg1 on p56lck Kinase and Cell Proliferation in Jurkat T Cells)

  • Hong, Hee-Youn;Na, Do-Seong;Kwon, Tae-Ik;Choi, Jung-Kap;Yoo, Gyurng-Soo
    • Journal of Ginseng Research
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    • 제19권2호
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    • pp.117-121
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    • 1995
  • We studied the effects of ginsenoside-$Rg_1$ (G-$Rg_1$) extracted from Panax ginseng C.A. Meyer on $p56^{kk}$ kinase and cell proliferation in Jurkat T cells. $p56^{kk}$ was maximally activated within 5 min after the treatment of 16.7 $\mu\textrm{g}$/ml of G-$Rg_1$ increasing the activity by 1.2-2 times relative to untreated control, thereafter its activity was gradually decreased to the level of untreated control. The action of EGTA on the kinase was altered by the addition of G-$Rg_1$, accompanying the band shift of $p56^{kk}$ to $p60^{kk}$. In addition, G-$Rg_1$promoted cell proliferation in a concentration-dependent manner. These results suggest that G-$Rg_1$ may be involved in T cell receptor-CD3 (TCR) signaling via the activation of $p56^{kk}$ and the chance of cellular calcium concentration.

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인체 급성백혈병 Jurkat T 세포에 있어서 L-canavanine에 의해 유도되는 세포자살기전에 미치는 단백질 티로신 키나아제 p56lck의 저해 효과 (A Natural L-Arginine Analog, L-Canavanine-Induced Apoptosis is Suppressed by Protein Tyrosine Kinase p56lck in Human Acute Leukemia Jurkat T Cells)

  • 박해선;전도연;우현주;류석우;김경민;김상국;박완;문병조;김영호
    • 생명과학회지
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    • 제19권11호
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    • pp.1529-1537
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    • 2009
  • L-arginine 구조유사체인 L-canavanine의 인체 급성백혈병 Jurkat T 세포에 대한 apoptosis 유도활성이 단백질 티로신키나아제 $p56^{lck}$에 어떻게 조절되는지를 규명하기 위해 $p56^{lck}$를 발현하는 Jurkat T 세포주 E6.1과 $p56^{lck}$-결손 Jurkat T 세포주 JCaM1.6에 있어서 L-canavanine의 세포독성, L-canavanine에 의해 유도되는 apoptotic DNA fragmentation 및 apoptotic sub-$G_1$ peak를 비교하여 본 바, $p56^{lck}$-negative JCaM1.6 세포가 $p56^{lck}$-positive E6.1 세포에 비해 L-canavanine의 apoptotis 유도활성에 훨씬 더 민감한 것으로 나타났다. 이러한 $p56^{lck}$-negative JCaM1.6 세포의 민감성은 JCaM1.6 세포에 $p56^{lck}$ 유전자를 transfection시켜 발현시키면 현저히 감소되었다. L-Canavanine에 의해 유도되는 apoptosis관련 현상들을 $p56^{lck}$-stable transfectant인 JCaM1.6/lck 세포와 empty vector-transfectant 인 $p56^{lck}$-negaive JCaM1.6/vector 세포에서 Western blot analysis로 비교한 결과, L-canavanine에 의해 유도되는 mitochondrial membrane potential (${\Delta\Psi}m$)의 감소, caspase-9, -8, -7 및 -3의 활성화, 그리고 PARP 및 $PLC{\gamma}$-1의 분해가 JCaM1.6/vector 세포에 비해 JCaM1.6/lck 세포에서 더 약하게 나타났다. JCaM1.6/lck 세포를 2.5 mM L-canavanine으로 처리한 다음 세포 내 $p56^{lck}$ kinase 활성의 변화를 $\alpha$-casein을 기질로 하여 시간 별로 측정한 결과, L-canavanine의 처리 후 15분만에 $p56^{lck}$ kinase의 활성이 약 1.6배 증가되었으며 이후 6시간 동안은 약 1.3~1.4 배정도 증가된 수준으로 kinase 활성이 유지되는 것으로 확인되었다. L-Canavanine에 의한 apoptosis의 개시에 Fas/FasL 상호작용이 관련되는지를 규명하기 위해 FADD-negative Jurkat T 세포주 I2.1, caspase-8-negative Jurkat T 세포주 I9.2 및 wild-type Jurkat T 세포주 A3에 대한 L-canavanine의 세포독성을 비교한 결과, A3와 I2.1 세포의 경우는 L-canavanine의 세포독성이 동일하게 나타났고, 특히 caspase-8가 결손된 I9.2 세포의 경우는 L-canavanine의 세포독성에 대한 민감성이 A3와 I2.1 세포에 비해 단지 미약하게만 완화되는 것으로 나타나, L-canavanine의한 apoptosis에는 Fas/FasL 상호작용이 관련되어 있지 않으며, 또한 caspase-8의 역할이 필수적이지 않음을 시사하였다. Jurkat T 세포에 있어서 L-canavanie에 의해 유도되는 sub-$G_1$ peak 및 caspases 활성화에 미치는 pan-caspase inhibitor (z-VAD-fmk), caspase-9 inhibitor (z-LEHD-fmk), caspase-3 inhibitor (z-DEVD-fmk), caspase-4 inhibitor (z-LEVD-fmk) 및 caspase-12 inhibitor (z-ATAD-fmk)의 영향을 조사한 결과, L-canavanie에 의한 apoptosis는 ${\Delta\Psi}m$의 감소, caspase-9 및 caspase -3의 활성화에 뒤따른 caspase-8 및 caspase-7의 활성화, 그리고 PARP의 분해의 순서로 유도되는 것으로 나타났으며, 아울러 caspase-9의 활성화와 함께 caspase-12의 활성화가 L-canavanine 처리에 따른 caspase-3의 활성화에 요구되는 것으로 확인되었다. 결론적으로, L-canavanine 처리에 의한 Jurkat T 세포의 apoptosis는 ${\Delta\Psi}m$ 감소, caspase-9, caspase-3 및 caspase-7의 활성화에 의해 유도되며, 이들 apoptosis 현상들은 $p56^{lck}$에 의해 negative regulation되었다.

C형 간염 바이러스 NS5B 단백질과 숙주의 p48 단백질의 상호작용에 의한 인터페론 저항성의 유도 (An Interferon Resistance Induced by the Interaction between HCV NS5B and Host p48)

  • 박소연;이종호;명희준
    • 한국미생물·생명공학회지
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    • 제36권4호
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    • pp.353-359
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    • 2008
  • Hepatitis C virus (HCV) is known as the causative agent of blood transmitted hepatitis. Two viral proteins, E2 and NS5A, are known to exert interferon resistance of HCV via PKR pathway. Here, we report a third protein, the RNA-dependent RNA polymerase (NS5B) of HCV, induced interferon resistance inhibiting p56 pathway. p56 was shown to interact with p48 subunit of eukaryotic initiation factor 3 (eIF3). This interaction inhibited formation of ternary complex in translation initiation. Using dual reporter assay system, we observed that the translation decreased when interferon alpha was added to the culture. But, in the presence of HCV NS5B, the translation partly recovered. NS5B and p48 subunit of eIF3 were shown to interact. This interaction seems to inhibit the interaction between p48 and p56. This is the first report that a virus exerts interferon resistance via p56 pathway.

Bacillus subtilis JK-56이 생산하는 chitinase isozyme의 정제와 특성 규명 (Purification and characterization of the chitinase from Bacillus subtilis JK-56)

  • 전홍기;김낙원;정영기
    • 생명과학회지
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    • 제12권1호
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    • pp.77-86
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    • 2002
  • 토양으로부터 chitinase를 생성하는 균주를 분리하여 동정한 결과 Bacillus subtilis로 판명되었으며, 분리한 균주를 Bacillus subtilis JK-56이라 명명하였다. B. subtilis JK-56의 chitinase 생산 최적 조건을 검토한 결과 1% chitin, 0.5% polypeptone, 0.1% KCI, 0.05% MnS $O_4$.4$H_2O$이며 초발 pH 7.0, 배양온도 37$^{\circ}C$에서 가장 많은 효소를 생산하였다. 본 균주가 생산하는 chitinase를 정제하기 위해서 native-PAGE를 이용해 효소활성 band를 확인한 결과, 1개의 강한 활성 band와 2개의 약한 활성 band를 가지는 isozyme으로 확인되었다. 확인된 isozyme을 정제한 결과, isozyme 중 1개의 강한 활성 band를 정제하였고 정제된 효소를 Chi-56A라고 명명하였다 Chi-56A의 효소 특성에 관해서 실험한 결과 분자량은 약 53kDa, pI는 4.3으로 확인되었다. 본 효소는 $65^{\circ}C$까지 상당히 안정하였으며 효소의 최대활성 온도도 $65^{\circ}C$로 확인되는 등 열에 대해 상당히 안정한 효소로 확인되었다. Collidal chitin에 대한 정제효소 Chi-56A의 $K_{m}$ 값은 17.33g/L였다. 그리고 pH 6.0에서 최대의 활성을 나타내었고, 산성범위보다 알칼리범위에서 안정한 것으로 나타났다. 또한 $Mn^{2+}$ 존재 하에서 높은 활성을 나타내었고 C $O^{2+}$$Mg^{2+}$ 존재 하에서도 활성이 약간 증가한 반면에 H $g^{2+}$ 존재 하에서는 상당한 저해를 받았다. Chito 올리고당에 대한 분해 산물을 HPLC로 확인해 본 결과 짝수개의 올리고당의 분해산물은 (GlcNAc)$_2$만을 생산하였고 홀수개의 올리고당에 대해서는 GlcNAc와 (GlcNAc)$_2$를 생산하는 것으로 비환원성 말단으로부터 이당체인 diacetyl chitobiose ((GlcNAc)$_2$)를 생산하는 exo형 chitinase로 추정 된다.

Cytotoxic activity and subset populations of peripheral blood natural killer cells in patients with chronic pain

  • Yoon, Jae Joon;Song, Ji A;Park, Sue Youn;Choi, Jeong Il
    • The Korean Journal of Pain
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    • 제31권1호
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    • pp.43-49
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    • 2018
  • Background: Chronic pain reportedly exerts complex effects on immune function. Natural killer (NK) cells are lymphocytes that play a critical role in cellular and innate immunity. This study examined changes in the subset populations and cytotoxic activity of peripheral blood NK cells in patients with chronic pain. Methods: Thirty patients with chronic moderate-to-severe pain (group P) and age-matched pain-free subjects (group NoP) were enrolled. Peripheral whole blood was analyzed for the percentage and expression of NK cell surface markers (CD56 and CD16) by flow cytometry. Cytotoxic activity was assayed by evaluating CD69 expression on $CD3^-/CD56^+NK$ cells. Results: The percentage of NK cells among total lymphocytes was not significantly different between groups P and NoP ($16.3{\pm}9.3$ vs. $20.2{\pm}10.5%$). Likewise, the percentages of two major NK cell subsets, $CD56^{bright}$ and $CD56^{dim}$, were also not significantly different between the two groups. However, the percentage of $CD56^{bright}/CD16^+$ subset, was slightly but significantly increased in group P ($1.0{\pm}0.9%$; P< 0.01) compared with group NoP ($0.5{\pm}0.6%$). The cytotoxicity of NK cells was not different between the two groups, showing similar CD69 expression (P vs. $NoP=29.2{\pm}15.2$ vs. $32.0{\pm}15.0%$). These findings were not influenced by pain intensity, opioid use, or disease causing pain in group P. Conclusions: NK cell cytotoxic activity and major subset populations, with the exception of an increased percentage of the $CD56^{bright}/CD16^+$ subset, are not significantly altered in patients with chronic severe pain.

Quantitative Structure-Activity Relationships and Molecular Docking Studies of P56 LCK Inhibitors

  • Bharatham, Nagakumar;Bharatham, Kavitha;Lee, Keun-Woo
    • Bulletin of the Korean Chemical Society
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    • 제27권2호
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    • pp.266-272
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    • 2006
  • Three-dimensional quantitative structure-activity relationship (3D-QSAR) models were developed for 67 molecules of 2-amino-benzothiazole-6-anilide derivatives against lymphocyte-specific protein tyrosine kinase (P56 LCK). The molecular field analysis (MFA) and receptor surface analysis (RSA) were employed for QSAR studies and the predictive ability of the model was validated by 15 test set molecules. Structure-based investigations using molecular docking simulation were performed with the crystal structure of P56 LCK. Good correlation between predicted fitness scores versus observed activities was demonstrated. The results suggested that the nature of substitutions at the 2-amino and 6-anilide positions were crucial in enhancing the activity, thereby providing new guidelines for the design of novel P56 LCK inhibitors.

중성자조사 금속 과망간산염의 반조효과 (Recoil Effects of Neutron-Irradiated Metal Permanganates)

  • Lee, Byung-Hun;Kim, Jung-Gwan
    • Nuclear Engineering and Technology
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    • 제20권2호
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    • pp.105-111
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    • 1988
  • 과망간산염들, 즉 과망간산 칼륨, 과망간산 암모늄, 과망간산 바륨에서 망간의 중성자 포획으로 야기되는 화학적 효과를 고찰하였다. $^{55}$Mn(n, r) $^{56}$ Mn 반응에서 생성된 방사성 망간 화학종, 즉 양이온 56/Mn, $^{56}$ MnO$_2$ 그리고 $^{56}$ MnO$_4$$^{-}$의 분포에 미치는 용제의 pH효과를 여러 가지 흡착제들과 이온교환체, 즉 제올이프 A-3, 카올리나이트, 알루미나, 이산화망간 그리고 도엑스 -50을 이용하여 고찰하였다. 카올리나이트와 알루미나에서 방사성 MnO$_4$$^{-}$의 분포가 대표적인 pH값인 4, 7 그리고 9 각각에서 다른흡착제와 이온교환체보다 높게 나타나며 동일한 흡착제일경우에는 pH 4 는 및 pH 9에서가 pH 7에서보다 높게 나타난다. $^{55}$Mn(n, r) $^{56}$ Mn 반응에 의하여 과망간산염에서 생성된 반조망간원자들의 열-어니어링 거동 또한 고찰하였다. 열-어니어링에서 $^{56}$ MnO$_4$$^{-}$의 잔류율은 10$0^{\circ}C$ 및 13$0^{\circ}C$ 처리에서 온도가 높아질수록 증가함을 보였다. 망간염의 반조효과는 hot zone model로 설명하였다.

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