• Title/Summary/Keyword: Small protein

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Cytogenetic Analysis of Bupleurum falcatum L. Cultivated in Korea (한국 재배종 시호의 세포유전학적 분석)

  • Chung, Sung-Hyun;Bang, Jae-Wook;Choi, Hae-Woon
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.61-65
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    • 1995
  • Karyotype analysis was carried out in four lines of Bupleurum falcatum L. cultivated in Korea and SDS-PAGE was applied to determine the seed protein profiles among the lines. Chromosomes were classified into two groups, large and small ones. Two kinds of karyotype, 2n=20 and 2n=26, were identified. Chromosome 1 of 2n=20 were all submedian, while that of 2n=26 were median. Chromosomes 2, 3 and 5 of 2n=20 showed polymorphism in size and arm-ratio. Chromosome 2 was submedian, while others were median in the line of 2n=26. Karyotypcs of cultivars native of Korea were similiar each other, while those introduced from Japan showed different patterns. In SDS PAGE gels, qualitative difference s in high molecular weight proteins, more than 45KD, were detected among the lines. The numbers of specific band were three in lines of 2n=20 and two in 2n=26.

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Studies on the mechanism of Nitric oxide (NO) induction in the Peritoneal Macrophage by HERBA SAURUI (HS) (삼백초(三白草)가 복강(腹腔) 대식세포(大食細胞)로부터 Nitric Oxide(NO) 유리기전(遊離機轉)에 대한 연구(硏究))

  • Jeon, Gil-Hwan;Shin, Min-Kyo;Song, Ho-Joon
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.36-49
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    • 1998
  • HERBA SAURURI (HS) has been known to use antiinflammatory drug. To investigated the mechanism of HS-induced NO synthesis, I evaluated the ability of protein kinase C (PKC) inhibitors such as staurosporine (STSN) or polyymyxin B to block HS-induced effects. HS alone had only a small effect, whereas in combination with $rIFN-{\gamma}$, markedly increased NO synthesis in a dose dependent manner. STSN and polymyxin B decreased NO synthesis, which had been induced by $rIFN-{\gamma}$, plus HS. Furthermore, prolonged incubation of the cells with phorbol ester, which down-regulates PKC activity abolished synergistic cooperative effect of HS with $rIFN-{\gamma}$ on NO synthesis. STSN and Polymyxin B potently inhibited HS-induced $TNF-{\alpha}$ secretion by $rIFN-{\gamma}$ plus HS. However, $rIFN-{\gamma}$ plus $TNF-{\alpha}-induced$ NO synthesis was not blocked by STSN or polymyxin B. On the other hand, tyrosine kinase inhibitor, genistein, blocked the NO synthesis and $TNF-{\alpha}$ secretion by $rIFN-{\gamma}$ plus HS. In conlusion, the present results strongly suggest that the capacity of HS to increase NO synthesis from $rIFN-{\gamma}-primed$ macrophages is the result of HS-induced $TNF-{\alpha}$ secretion via the signal transduction pathway of PKC and tyrosine kinase.

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Inhibitory Effect of Immunoglobulin E Production by Poncirus tripoliata (지실(枳實)에 의한 면역(免疫)글로블린 E 생성(生成)의 억제효과(抑制效果))

  • Kim, Hyeong-Kyun;Kweon, Yong-Taek
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.419-429
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    • 1998
  • Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) has been used for the treatment of allergic disease. IgE is normally one of the least abundant immunoglobulin (Ig) isotypes in the serum of both humans and several species of experimental animals: however a number of different stimuli can result in profound increases in IgE levels relative to other isotypes. In rodents, infection with many parasitic helminths can cause approximately 100-fold elevation in IgE within 2 wks. Immunization of mice with small amounts of protein antigens on alum also results in 10-fold to fold increase in total serum IgE, much of it specific for the immunizing antigen. In this experiment, I investigated the effect of an aqueous extract of Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) on a in vivo and in vitro IgE production. PTFE dose-dependently inhibited the serum levels of IgE induced by antigens. The regulation of IgE synthesis is influenced by T cells and T cell derived factors. IL -4, a T cell-derived cytokine, has been shown to stimulate murine IgE synthesis both in vitro and in vivo. Current evidence suggests that IL-4 induces IgE synthesis in the mouse by stimulating H chain isotype switch. Lipopolysaccharide (LPS) plus IL-4 cause about l00-fold increase in IgE secretion by murine B cells. The effects of PTFE on the IL-4-dependent IgE response by mouse whole spleen cells were studied. Whole spleen cells were cultured for 7 days in the presence of LPS plus IL-4 and PTFE and the supernatants were assayed for IgE. IL-4 dependent IgE production of LPS-stimulated whole spleen cells was inhibited by PTFE. Moreover, in the present study using U266Bl human IgE-bearing B cells, I found that PTFE inhibited the production of IgE activated by LPS plus IL-4. These results indicate that PTFE have antiallergic activity by inhibition of IgE production from B cells.

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Partial Purification and Characterization of Minor Form of Phosphofructokinase from the Host Fraction of Chickpea(Cicer arietinum L. cv. Amethyst) Nodules (병아리콩(Cicer arietinum L. cv. Amethyst) 근류내의 플라스티드 포스포프룩토오스 키나아제의 분리 및 특성)

  • Lee, Hoi-Seon
    • Applied Biological Chemistry
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    • v.41 no.5
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    • pp.355-362
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    • 1998
  • The minor form of phosphofructokinase (EC 2.7.1.11; PFK), which was suggested to be of plastid origin from the host fraction of chickpea nodules, was isolated as a small protein with apparent molecular mass near 220 kDa and purified to a high degree. SDS-PAGE and western blot indicated that the enzyme was made up of a homotetrameric structure (55 kDa). The enzyme had sharp pH profiles with maximal activities at pH 8 and displayed Michaelis-Menten kinetics with respect to Fru-6-P and nucleoside triphosphate substrate at the pH optimum (pH 8) and at pH 7. MgATP was the most effective phosphoryl donor. Phosphoenolpyruvate was a potent inhibitor of minor PFK activity, and the enzyme was also strongly inhibited by 3-phosphoglycerate, 2-phosphoglycerate, and to a lesser extent, PPi. Minor PFK was weakly activated by KCl, NaCl and Pi, and was inhibitory at high concentration of KCl and Pi.

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Streptochlorin Isolated from Streptomyces sp. Induces Apoptosis in Human Hepatocarcinoma Cells Through a Reactive Oxygen Species-Mediated Mitochondrial Pathway

  • Shin, Dong-Yeok;Shin, Hee-Jae;Kim, Gi-Young;Cheong, Jae-Hun;Choi, Il-Whan;Kim, Se-Kwon;Moon, Sung-Kwon;Kang, Ho-Sung;Choi, Yung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1862-1867
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    • 2008
  • Streptochlorin is a small molecule isolated from marine Streptomyces sp. that is known to have antiangiogenic and anticancer properties. In this study, we examined the effects of this compound on reactive oxygen species (ROS) production and the association of these effects with apoptotic tumor cell death, using a human hepatocarcinoma Hep3B cell line. The results of this study demonstrated that streptochlorin mediates ROS production, and that this mediation is followed by a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\Psi}_m$), activation of caspase-3, and downregulation of antiapoptotic Bcl-2 protein. The quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the streptochlorin-induced apoptosis effects via inhibition of ROS production, MMP collapse, and the subsequent activation of caspase-3. These observations clearly indicate that ROS are involved in the early molecular events in the streptochlorin-induced apoptotic pathway. Taken together, our data imply that streptochlorin-induced ROS is a key mediator of MMP collapse, which leads to the caspase-3 activation, culminating in apoptosis.

Effects of the Feed and Probiotic Feeding on the Improvement of Hoggery Environment and the Productivity of Swine (사료 및 보조사료로서의 생균제 급여에 따른 돈사 환경개선과 돼지 생산성에 미치는 영향)

  • Lee, Enu-Young;Lim, Joung-Soo
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.200-209
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    • 2011
  • Animals produce important co-products such as meat, milk, and egg. Higher consumption and urbanization asked for more animal products and the demand was so strong that lager livestocks are now being raised densely in small farm. Large production of excreta and maldor is an inevitable consequence of condensed breeding. If this malodor couldn't be controlled, it could be chief obstacle to development of both livestock industry and environment of future. Major odor produced from livestock environments could be subdivided into four major sections: volatile fatty acids, ammonia and volatile amine, indole and phenols, and sulfur compounds. More than half of nitrogen excreted urea, so low protein feeding, synthetic amino acid feeding and supplementing with digestive enzyme, microbial agents and/or probiotics are methods for reducing nitrogen excretion. A lot of studies about feeding and probiotics, co-feed have been researched to improve environment and/or productivity in livestock industry.

Enzymatic Characteristics of a Highly Thermostable β-(1-4)-Glucanase from Fervidobacterium islandicum AW-1 (KCTC 4680)

  • Jeong, Woo Soo;Seo, Dong Ho;Jung, Jong Hyun;Jung, Dong Hyun;Lee, Dong-Woo;Park, Young-Seo;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.271-276
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    • 2017
  • A highly thermostable ${\beta}-(1-4)-glucanase$ (NA23_08975) gene (fig) from Fervidobacterium islandicum AW-1, a native-feather degrading thermophilic eubacterium, was cloned and expressed in Escherichia coli. The recombinant FiG (rFiG) protein showed strong activity toward ${\beta}-{\small{D}}-glucan$ from barley (367.0 IU/mg), galactomannan (174.0 IU/mg), and 4-nitrophenyl-cellobioside (66.1 IU/mg), but relatively weak activity was observed with hydroxyethyl cellulose (5.3 IU/mg), carboxymethyl cellulose (2.4 IU/mg), and xylan from oat spelt (1.4 IU/mg). rFiG exhibited optimal activity at $90^{\circ}C$ and pH 5.0. In addition, this enzyme was extremely thermostable, showing a half-life of 113 h at $85^{\circ}C$. These results indicate that rFiG could be used for hydrolysis of cellulosic and hemicellulosic biomass substrates for biofuel production.

Regulation of AKT Activity by Inhibition of the Pleckstrin Homology Domain-PtdIns(3,4,5)P3 Interaction Using Flavonoids

  • Kang, Yerin;Jang, Geupil;Ahn, Seunghyun;Lee, Youngshim;Shin, Soon Young;Yoon, Youngdae
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1401-1411
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    • 2018
  • The serine-threonine kinase AKT plays a pivotal role in tumor progression and is frequently overactivated in cancer cells; this protein is therefore a critical therapeutic target for cancer intervention. We aimed to identify small molecule inhibitors of the pleckstrin homology (PH) domain of AKT to disrupt binding of phosphatidylinositol-3,4,5-trisphosphate (PIP3), thereby downregulating AKT activity. Liposome pulldown assays coupled with fluorescence spectrometry were used to screen flavonoids for inhibition of the AKT PH-PIP3 interaction. Western blotting was used to determine the effects of the inhibitors on AKT activation in cancer cells, and in silico docking was used for structural analysis and optimization of inhibitor structure. Several flavonoids showing up to 50% inhibition of the AKT PH-PIP3 interaction decreased the level of AKT activation at the cellular level. In addition, the modified flavonoid showed increased inhibitory effects and the approach would be applied to develop anticancer drug candidates. In this study, we provide a rationale for targeting the lipid-binding domain of AKT, rather than the catalytic kinase domain, in anticancer drug development.

XENOTRANSPLANT OF HUMAN BONE MARROW STROMAL CELLS; EFFECT ON THE REGENERATION OF AXOTOMIZED INFRAORBITAL NERVE IN RATS (인간 골수 기질세포 이종이식이 백서의 축삭절단 안와하 신경 재생에 미치는 효과)

  • Park, Eun-Jin;Kim, Eun-Seok;Kim, Jin-Man;Kim, Hyun-Ok;Yum, Kwang-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.239-247
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    • 2005
  • This study demonstrated that xenogenic human marrow mesenchymal stem cells (hMSCs) could elicit the regeneration of the sensory nerve after axotomy in the adult rats infraorbital nerves without immunosuppression. For this, we evaluated the behavioral testing for functional recovery of the nerve and histological findings at weeks 3 and 5 compared to controls. Xenogenic hMSCs did not evoke any significant inflammatory or immunologic reaction after systemic and local administrations. HMSCs-treated rats exhibited significant improvement on sensory recovery tested with von Frey monofilaments. At 5 postoperative weeks, in the hMSCs treated nerve, expression of myelin basic protein (MBP), neurofilament (NF) at the site of axotomy was higher than control. And mRNA expression of neurotropin receptor Trk precursor (TrkPre), nerve growth factor receptor (NGFR) and neuropeptide (NPY) in trigeminal ganglion were also higher. The number of myelinated nerve at distal stump and cells in trigeminal ganglion were higher in hMSC treated rats. So it was supposed that transplanted MSCs contributed to reducing post-traumatic degeneration and production of neurotrophic factors. Immunofluorescence labeling showed small portion of hMSCs (<10%) expressed a phenotypic marker of Schwann cell (S-100). Xenogenic or allogenic mesenchymal stem cells might have immune privileged characteristics and useful tool for cell based nerve repair.

Inhibition of p90RSK activation sensitizes triple-negative breast cancer cells to cisplatin by inhibiting proliferation, migration and EMT

  • Jin, Yujin;Huynh, Diem Thi Ngoc;Kang, Keon Wook;Myung, Chang-Seon;Heo, Kyung-Sun
    • BMB Reports
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    • v.52 no.12
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    • pp.706-711
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    • 2019
  • Cisplatin (Cis-DDP) is one of the most widely used anti-cancer drugs. It is applicable to many types of cancer, including lung, bladder, and breast cancer. However, its use is now limited because of drug resistance. p90 ribosomal S6 kinase (p90RSK) is one of the downstream effectors in the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) pathway and high expression of p90RSK is observed in human breast cancer tissues. Therefore, we investigated the role of p90RSK in the Cis-DDP resistance-related signaling pathway and epithelial-mesenchymal transition (EMT) in breast cancer cells. First, we discovered that MDA-MB-231 cells exhibited more Cis-DDP resistance than other breast cancer cells, including MCF-7 and BT549 cells. Cis-DDP increased p90RSK activation, whereas the inactivation of p90RSK using a small interfering RNA (siRNA) or dominant-negative kinase mutant plasmid overexpression significantly reduced Cis-DDP-induced cell proliferation and migration via the inhibition of matrix metallopeptidase (MMP)2 and MMP9 in MDA-MB-231 cells. In addition, p90RSK activation was involved in EMT via the upregulation of mRNA expression, including that of Snail, Twist, ZEB1, N-cadherin, and vimentin. We also investigated NF-κB, the upstream regulator of EMT markers, and discovered that Cis-DDP treatment led to NF-κB translocation in the nucleus as well as its promoter activity. Our results suggest that targeting p90RSK would be a good strategy to increase Cis-DDP sensitivity in triple-negative breast cancers.