• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.355-362
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• 2000

The Herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) gene in the pHLA-21 plasmid was inserted into a baculovirus (Hyphantria cunea nuclear polyhedrosis virus) expression vector (lacZ-HcNPV) to construct a recombinant virus gB-HcNPV expressing gB. Spodoptera frugiperda cells infected with this recombinant virus synthesized and processed gB of approximately 120 kDa, which cross-reacted with the monoclonal antibody to gB. The recombinant gB was identified on the membrane of the insect cells using an immunofluorescence assay. Antibodies to this recombinant raised in mice recognize the viral gB and neutralized the infectivity of the HSV-1 in vitro. These results show that the gB gene has the potential to be expressed in insect cells. They also demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the lacZ-HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• BMB Reports
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• 제41권10호
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• pp.705-709
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• 2008

NF-${\kappa}B$ is a transcriptional regulator involved in many biological processes including proliferation, survival, and differentiation. Recently, we reported that expression and activity of NF-${\kappa}B$ is comparatively low in undifferentiated human embryonic stem (ES) cells, but increases during differentiation. Here, we found a lower expression of NF-${\kappa}B$ p65 protein in mouse ES cells when compared with mouse embryonic fibroblast cells. Protein levels of NF-${\kappa}B$ p65 and relB were clearly enhanced during retinoic acid-induced differentiation. Furthermore, increased DNA binding activity of NF-${\kappa}B$ in response to TNF-$\alpha$, an agonist of NF-${\kappa}B$ signaling, was seen in differentiated but not undifferentiated mouse ES cells. Taken together with our previous data in human ES cells, it is likely that NF-${\kappa}B$ expression and activity of the NF-${\kappa}B$ signaling pathway is comparatively low in undifferentiated ES cells, but increases during differentiation of ES cells in general.

• Biomolecules & Therapeutics
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• 제21권6호
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• pp.435-441
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• 2013

Emodin, a naturally occurring anthraquinone derivative isolated from Polygoni cuspidati radix, has several beneficial pharmacologic effects, which include anti-cancer, anti-diabetic, and anti-inflammatory activities. In this study, the authors examined the effect of emodin on the production of proinflammatory cytokines, such as, tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, in mouse bone marrow-derived mast cells (BMMCs) stimulated with phorbol 12-myristate 13-acetate (PMA) plus the calcium ionophore A23187. To investigate the mechanism responsible for the regulation of pro-inflammatory cytokine production by emodin, the authors assessed its effects on the activations of transcriptional factor nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and mitogen-activated protein kinases (MAPKs). Emodin attenuated the nuclear translocation of (NF)-${\kappa}B$ p65 and its DNA-binding activity by reducing the phosphorylation and degradation of $I{\kappa}B{\alpha}$ and the phosphorylation of $I{\kappa}B$ kinase B (IKK). Furthermore, emodin dose-dependently attenuated the phosphorylations of MAPKs, such as, extracellular signal-regulated kinase 1/2 (ERK1/2), p38 MAP kinase, and the stress-activated protein kinases (SAPK)/c-Jun-N-terminal kinase (JNK). Taken together, the findings of this study suggest that the anti-inflammatory effects of emodin on PMA plus A23187-stimulated BMMCs are mediated via the inhibition of NF-${\kappa}B$ activation and of the MAPK pathway.

• 한국식품과학회지
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• 제31권1호
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• pp.219-223
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• 1999

우리나라의 전통 발효식품인 김치에서 혈전용해 효소를 생산하는 미생물을 분리하고 그 중 Bacillus amyloliquefaciens, Bacillus brevis, Micrococcus luteus의 세 종을 Bergey's manual of systematic bacteriology등에 의하여 동정하였다. 분리된 미생물을 효소 유도 배지에서 배양한 결과 B. amyloliquefaciens는 2.58 plasmin unit/mL, B. brevis는 1.48 plasmin unit/mL, 그리고 M. luteus는 2.03 plasmin unit/mL의 혈전용해효소 생산능을 보여 주였다. 각 균주에서 생산된 세포의 단백질을 SDS-PAGE와 fibrin zymography assay에 의해 분석한 결과 B. brevis와 M. luteus에서는 서로 다른 분자량을 가진 $3{\sim}4$개의 혈전용해 효소가 존재하였으며 B. amyloliquefaciens에서는 분자량이 약 29 kDa인 단일 band의 혈전용해 효소가 생산되었음을 확인하였다.

• Parasites, Hosts and Diseases
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• 제29권1호
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• pp.1-8
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• 1991

스파르가눔 생리식염수 추출액 내에 포함되어 있는 성분단백질 중 스파르가눔증 환자 혈청내 특이 IgG항체와 민감하고 특이하게 반응하는 항원단벼질인 36, 29 kDa단백질을 단세포군 항체를 이용한 면역친화성 크로마토그 래피로 순수분리할 수 있음은 이미 보고하였다. 이 연구에서는 스파르가눔 추출액 내에 포함된 이 36, 29 kDa단백질이 젤라틴을 고리로 한 친화성 크로마토그래피로 훨씬 쉽게 순수분리할 수 있음을 증명하고자 하였다. 젤라틴을 고리로 부착시킨 Sepharose 4B column에 스파르가눔 추출액을 통과시키고 젤라틴에 부착한 단백질은 4 M urea/0.1M NaCl 용액을 분리완충액으로 분리하였다. 이렇게 분리한 단백질은 SDS-PAGE에서 36, 29 kDa band로 구성되어 있었고, SDS-PAGE/immunoblot 결과 환자의 polyclonal 항체는 이들 band에만 반응하였다. 스파르가눔증, 기타 기생충증 환자 및 건강대조군 혈청내 스파르가눔 특이항체가(IgG)를 면역효소측정 법으로 측정 한 결과 순수분리한 이 단백질은 특히 특이도가 95.8%로 생리식염수 추출액의 89%보다 우수하였고 민감도는 차이가 없었다. 이상의 결과는 젤라틴을 고리로 이용한 친화성 크로마토그래피는 스파르가눔 생리식염수 추출액 내의 36 및 29 kDa 단백질을 간편하게 순수분리할 수 있고 단백질의 항원성도 유지할 수 있음을 보이고 있었다.

• 생약학회지
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• 제45권3호
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• pp.262-267
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• 2014

Royal jelly composes of many components, especially protein. Protein is a major factor which cause allergy. We focused on water soluble royal jelly (WSRJ) that was removed allergy - inducing protein. 10-hyroxy-2-decenoic acid content of WSRJ is 2.42 g/100 g, which is double compared to that of lypophilized RJ. To further access WSRJ as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that WSRJ increased the cell viability in B16F1 melanoma cell and WSRJ (1~10 mg/ml) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. WSRJ inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that WSRJ induces the down regulation of melanogenesis by inhibiting tyrosinase activation.

• 생명과학회지
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• 제22권9호
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• pp.1137-1144
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• 2012

염증반응은 유해한 물질이나 병원체에 대항하여 활성화되는 생체 방어 기전이다. 그러나 과도한 염증반응은 그 자체가 생체에 좋지 않은 영향을 미칠 수 있다. 대식세포는 지질다당류와 같은 병원체를 인식한 후, NF-${\kappa}B$ 경로의 활성화를 포함한 다양한 경로를 통하여 산화질소와 같은 염증매개인자들을 분비하는 면역세포이다. 본 연구에서는 지질다당류로 활성화시킨 RAW 264.7 대식세포를 이용하여 등골나물(Eupatorium chinensis var. simplicifolium) 뿌리, 줄기 그리고 꽃 추출물들의 항염증 효과를 알아보았다. 그 중 등골나물 뿌리의 추출물은 농도의존적으로 산화질소의 생성을 감소시켰으며, 산화질소 합성유도효소(inducible nitric oxide synthase)와 고리형 산소화효소-2(cyclooxygenase-2)의 발현을 통계적으로 유의하게 감소시켰다. 또한 등골나물 뿌리의 추출물은 NF-${\kappa}B$ 경로에 있는 MAP (mitogen activated protein) 인산화효소와 단백질 인산화효소 B (protein kinase B)의 활성화를 감소시켰으며, 억제적 kappa B (inhibitory kappa B)의 분해 또한 감소시키는 것을 관찰하였다. 이러한 결과는 등골나물 뿌리의 추출물이 NF-${\kappa}B$ 경로와 산화질소 합성유도효소 발현의 억제를 통하여 항염증작용을 나타낼 수 있음을 제시한다.

• 약학회지
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• 제42권5호
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• pp.487-493
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• 1998

In the previous study we described the antitumor effect of GLB, a protein-polysaccharide fraction separated from the growing tips of Ganoderma lucidum, against sarcoma 18 0 solid tumor in ICR mice. In this study, we separated an acidic protein-polysaccharide fraction, GLB-A, and a basic protein-polyaccharide fraction, GLB-B, from GLB by differential precipitation, and elucidated their antitumor and immunomodulatory activities. When ip injected at the dose of 50mg/kg/day into the ICR mice, GLB-A and GLB-B inhibited the growth of ip implantated sarcoma 180 cells by 32.4% and 21.0%, respectively. Of these, GLB-A increased the % lymphoblast in the spleen of the tumor-bearing and the normal mice by 20.9% and 123.0%, and the CD4/CD8 ratio by 73.3% and 22.4%, respectively. GLB-A also increased the expression of CD25 (IL-2 receptor alpha ch0ain) in normal mice by 82.0%. These results strongly suggest that GLB-A is a promising candidate for antitumor immunomodulatory medicine.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.241-242
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• 2003

The effects of N-acetylphytospingosine(NAPS), one of the phytospingosine derivatives, on melanogenesis of B 16F 1 0 mouse melanoma cell lines were investigated. We assessed the effect of NAPS on the depigmentation of B16F10 cells. The melanin content of cells was significantly reduced by NAPS. We examined the inhibitory effect of NAPS on tyrosinase activity using L-dopa as a substrate and the results showed that tyrosinase activity was inhibited in a does-dependent manner. The mRNA level of tyrosinase as well as that of tyrosinase related protein-l (TRP-l) and tyrosinase related protein-2 (TRP-2) genes were not affected by NAPS based on a reverse transcription-polymerase chain reaction (RT-PCR) assay. We also performed a Western blotting analysis using anti-tyrosinase antibody. It showed that there is no change in tyrosinase protein level after treatment of NAPS. These results suggest that the depigmenting mechanism of NAPS in B16F10 melanoma cells involves inhibition of melanosomal tyrosinase activity, rather than the mRNA expression or protein level of tyrosinase.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.81.2-82
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• 2003

Plants have evolved along with pathogens, and they have developed sophisticated defense systems against specific microorganisms to survive. G-protons are considered one of the upstream signaling components working as a key for the defense signal transduction pathway. For activation and inactivation of G-protein, GTP-biding proteins are involved. GTP -binding proteins are found in all organisms. Small GTP-binding proteins, having masses of 21 to 30kD, belong to a superfamily, often named the Ras supefamily because the founding members are encoded by human Ras genes initially discovered as cellular homologs of the viral ras oncogene. Members of this supefamily share several common structural features, including several guanine nucleotide binding domains and an effector binding domain. However, exhibiting a remarkable diversity in both structure and function. They are important molecular switches that cycle between the GDP-bound inactive form into the GTP-bound active form through GDP/GTP replacement. In addition, most GTP-binding proteins cycle between membrane-bound and cytosolic forms. such as the RAC family are cytosolic signal transduction proteins that often are involved in processing of extracellular stimuli. Plant RAC proteins are implicated in regulation of plant cell architecture secondary wall formation, meristem signaling, and defense against pathogens. But their molecular mechanisms and functions are not well known. We isolated a RacB homolog from rice to study its role of defense against pathogens. We introduced the constitutively active and the dominant negative forms of the GTP-hinging protein OsRacB into the wild type rice. The dominant negative foms are using two forms (full-sequence and specific RNA interference with RacB). Employing southern, and protein analysis, we examine to different things between the wild type and the transformed plant. And analyzing biolistic bombardment of onion epidermal cell with GFP-RacB fusion protein revealed association with the nucle.