• Title/Summary/Keyword: $\alpha$-CD

Search Result 678, Processing Time 0.027 seconds

Purification and Properties of Extracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • Yu, Tae-Shick;Kim, Tae-Hyun
    • Journal of Microbiology and Biotechnology
    • /
    • v.9 no.2
    • /
    • pp.173-178
    • /
    • 1999
  • The extracellular cytosine deaminase (EC 3.5.4.1) from Chromobacterium violaceum YK 391 was purified 264.7-fold with an overall yield of 14.3%. The enzyme was for the first time homogeneous by the criteria of polyacrylamide gel electrophoresis performed in the absence and in the presence of sodium dodecyl sulfate. The molecular weight of the purified enzyme was estimated to be about 156 kDa. The enzyme consisted of two identical subunits of approximate molecular weight 78 kDa. The isoelectric point of the enzyme was pH 5.55. The enzyme had a pH optimum of 7.5 and a temperature optimum of around 40 to $45^{\circ}C$. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 5-methylcytosine, and 6-azacytosine, but not 5-azacytosine. The extracellular cytosine deaminase is believed to be unique because it was active not only on cytosine but also on cytidine. The apparent $K_m$ values for cytosine, 5-fluorocytosine, cytidine, and 5-methylcytosine were determined to be 1.55 mM, 5.52 mM, 10.4 mM, and 67.2 mM, respectively. The enzyme activity was strongly inhibited by heavy metal ions such as $Fe^{2+},Pb^{2+},Cd^{2+},Zn^{2+}, Hg^{2+}, and Cu^{2+}$ at 1 mM, and completely by $\alpha,\alpha$'-dipyridyl, and $\rho$-chloromercuribenzoate at 1 mM, and weakly inhibited by 1mM ο-phenanthroline. The enzyme activity was not affected by various nucleosides and nucleotides.

  • PDF

Probing α/β Balances in Modified Amber Force Fields from a Molecular Dynamics Study on a ββα Model Protein (1FSD)

  • Yang, Changwon;Kim, Eunae;Pak, Youngshang
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.6
    • /
    • pp.1713-1719
    • /
    • 2014
  • 1FSD is a 28-residue designed protein with a ${\beta}{\beta}{\alpha}$ motif. Since this protein displays most essential features of protein structures in such a small size, this model protein can be an outstanding system for evaluating the balance in the propensity of the secondary structures and the quality of all-atom protein force fields. Particularly, this protein would be difficult to fold to its correct native structure without establishing proper balances between the secondary structure elements in all-atom energy functions. In this work, a series of the recently optimized five amber protein force fields [$ff03^*$, $f99sb^*$-ildn, ff99sb-${\phi}^{\prime}$-ildn, ff99sb-nmr1-ildn, ff99sb-${\Phi}{\Psi}$(G24, CS)-ildn] were investigated for the simulations of 1FSD using a conventional molecular dynamics (MD) and a biased-exchange meta-dynamics (BEMD) methods. Among those tested force fields, we found that ff99sb-nmr1-ildn and ff99sb-${\Phi}{\Psi}$(G24, CS)-ildn are promising in that both force fields can locate the native state of 1FSD with a high accuracy (backbone rmsd ${\leq}1.7{\AA}$) in the global free energy minimum basin with a reasonable energetics conforming to a previous circular dichroism (CD) experiment. Furthermore, both force fields led to a common set of two distinct folding pathways with a heterogeneous nature of the transition state to the folding. We anticipate that these force fields are reasonably well balanced, thereby transferable to many other protein folds.

Structure-activity relationships of the intramolecular disulfide bonds in coprisin, a defensin from the dung beetle

  • Lee, Jaeho;Lee, Daeun;Choi, Hyemin;Kim, Ha Hyung;Kim, Ho;Hwang, Jae Sam;Lee, Dong Gun;Kim, Jae Il
    • BMB Reports
    • /
    • v.47 no.11
    • /
    • pp.625-630
    • /
    • 2014
  • Defensins, which are small cationic molecules produced by organisms as part of their innate immune response, share a common structural scaffold that is stabilized by three disulfide bridges. Coprisin is a 43-amino acid defensin-like peptide from Copris tripartitus. Here, we report the intramolecular disulfide connectivity of cysteine-rich coprisin, and show that it is the same as in other insect defensins. The disulfide bond pairings of coprisin were determined by combining the enzymatic cleavage and mass analysis. We found that the loss of any single disulfide bond in coprisin eliminated all antibacterial, but not antifungal, activity. Circular dichroism (CD) analysis showed that two disulfide bonds, Cys20-Cys39 and Cys24-Cys41, stabilize coprisin's ${\alpha}$-helical region. Moreover, a BLAST search against UniProtKB database revealed that coprisin's ${\alpha}$-helical region is highly homologous to those of other insect defensins.

Ginsenoside Rd alleviates mouse acute renal ischemia/reperfusion injury by modulating macrophage phenotype

  • Ren, Kaixi;Jin, Chao;Ma, Pengfei;Ren, Qinyou;Jia, Zhansheng;Zhu, Daocheng
    • Journal of Ginseng Research
    • /
    • v.40 no.2
    • /
    • pp.196-202
    • /
    • 2016
  • Background: Ginsenoside Rd (GSRd), a main component of the root of Panax ginseng, exhibits anti-inflammation functions and decreases infarct size in many injuries and ischemia diseases such as focal cerebral ischemia. M1 Macrophages are regarded as one of the key inflammatory cells having functions for disease progression. Methods: To investigate the effect of GSRd on renal ischemia/reperfusion injury (IRI) and macrophage functional status, and their regulatory role on mouse polarized macrophages in vitro, GSRd (10-100 mg/kg) and vehicle were applied to mice 30 min before renal IRI modeling. Renal functions were reflected by blood serum creatinine and blood urea nitrogen level and histopathological examination. M1 polarized macrophages infiltration was identified by flow cytometry analysis and immunofluorescence staining with $CD11b^+$, $iNOS^+$/interleukin-12/tumor necrosis factor-${\alpha}$ labeling. For the in vitro study, GSRd ($10-100{\mu}g/mL$) and vehicle were added in the culture medium of M1 macrophages to assess their regulatory function on polarization phenotype. Results: In vivo data showed a protective role of GSRd at 50 mg/kg on Day 3. Serum level of serum creatinine and blood urea nitrogen significantly dropped compared with other groups. Reduced renal tissue damage and M1 macrophage infiltration showed on hematoxylin-eosin staining and flow cytometry and immunofluorescence staining confirmed this improvement. With GSRd administration, in vitro cultured M1 macrophages secreted less inflammatory cytokines such as interleukin-12 and tumor necrosis factor-${\alpha}$. Furthermore, macrophage polarization-related pancake-like morphology gradually changed along with increasing concentration of GSRd in the medium. Conclusion: These findings demonstrate that GSRd possess a protective function against renal ischemia/reperfusion injury via downregulating M1 macrophage polarization.

Development of the Near Infrared Camera System for Astronomical Application

  • Moon, Bong-Kon
    • The Bulletin of The Korean Astronomical Society
    • /
    • v.35 no.1
    • /
    • pp.39.2-39.2
    • /
    • 2010
  • In this paper, I present the domestic development of near infrared camera systems for the ground telescope and the space satellite. These systems are the first infrared instruments made for astronomical observation in Korea. KASINICS (KASI Near Infrared Camera System) was developed to be installed on the 1.8m telescope of the Bohyunsan Optical Astronomy Observatory (BOAO) in Korea. KASINICS is equipped with a $512{\times}512$ InSb array enable L band observations as well as J, H, and Ks bands. The field-of-view of the array is $3.3'{\times}3.3'$ with a resolution of 0.39"/pixel. It employs an Offner relay optical system providing a cold stop to eliminate thermal background emission from the telescope structures. From the test observation, limiting magnitudes are J=17.6, H=17.5, Ks=16.1 and L(narrow)=10.0 mag at a signal-to-noise ratio of 10 in an integration time of 100 s. MIRIS (Multi-purpose InfraRed Imaging System) is the main payload of the STSAT-3 in Korea. MIRIS Space Observation Camera (SOC) covers the observation wavelength from $0.9{\mu}m$ to $2.0{\mu}m$ with a wide field of view $3.67^{\circ}{\times}3.67^{\circ}$. The PICNIC HgCdTe detector in a cold box is cooled down below 100K by a micro Stirling cooler of which cooling capacity is 220mW at 77K. MIRIS SOC adopts passive cooling technique to chill the telescope below 200K by pointing to the deep space (3K). The cooling mechanism employs a radiator, a Winston cone baffle, a thermal shield, MLI of 30 layers, and GFRP pipe support in the system. Opto-mechanical analysis was made in order to estimate and compensate possible stresses from the thermal contraction of mounting parts at cryogenic temperatures. Finite Element Analysis (FEA) of mechanical structure was also conducted to ensure safety and stability in launching environments and in orbit. MIRIS SOC will mainly perform the Galactic plane survey with narrow band filters (Pa $\alpha$ and Pa $\alpha$ continuum) and CIB (Cosmic Infrared Background) observation with wide band filters (I and H) driven by a cryogenic stepping motor.

  • PDF

EFFECTS OF CYTOKINES ON THE CELL PROLIFERATION OF CYTOLYTIC T CELL LINE CTLL - 2 (Cytolytic T cell line CTLL - 2의 세포증식에 미치는 cytokine의 효과)

  • Seo, Yang-Ja;Lee, In-Kyu;Lee, Jin-Young;Oh, Kwi-Ok;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
    • /
    • v.23 no.3
    • /
    • pp.454-460
    • /
    • 1993
  • Abnormalities of the T cell subsets have been detected in the immunologically mediated disease sites such as periodontal lesions which are attributable to the regulatory effect of cell differentiation and specific chemokinetic effect of various cytokines. Macrophage Inflammatory protein$(MIP)-1{\alpha}$ and gammain terferon$({\gamma}-IFN)$ serve as important immunoregulatory molecules through which growth and differentiation of specific T cell subsets are known to be negatively regulated. Murine cytolytic T cell line CTLL-2 were used to perform the [$^3H$]-thymidine incorporation test, by which we obtained more comprehensive view in regulatory actions of cytokines on the T cell subset proliferation. 1. $rMIP-{\alpha}$(200ng/ml) and $r{\gamma}-IFN$(100U/ml) appreared to suppress the proliferation rate to CTLL-2 by 74 and 86% respectively, and the suppressive action of two cytokines were synergisic. 2. Culture supernatant of anti-CD3 mAb-stimulated mouse splenocyte enhanced the proliferation rate of CTLL-2 up to 10-fold with dose-dependent manner. However, culture supernatant of unstimulated splenocyte showed only 2-fold increase in the proliferation rate. 3. CTLL-2 cell proliferation was strictly IL-2 dependent.

  • PDF

Immunological Synergistic Effects of Combined Treatment with Herbal Preparation (HemoHIM) and Red Ginseng Extracts (마우스세포를 이용한 홍삼추출물과 생약복합추출물의 병용 처리에 따른 면역활성 효과)

  • Byun, Myung-Woo;Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.44 no.2
    • /
    • pp.182-190
    • /
    • 2015
  • This present study demonstrates the immunological synergistic effects of herbal preparation (HemoHIM) and red ginseng powder granule in various immune cell models (bone marrow-derived macrophages, dendritic cells, and mouse splenocytes) from mice. Both herbal preparation and red ginseng extracts were treated to bone-marrow derived macrophages, dendritic cells, and mouse splenocytes, and there was no cytotoxicity at a dose below $200{\mu}g/mL$. Cell proliferation and cytokine [tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12] production tested in bone marrow-derived macrophages and dendritic cells significantly increased upon combined treatment. Cell surface marker (CD 80/86, MHC class I/II)-mediated immune cell activation was highly elevated by combined treatment. For cytokine production in splenocytes, combined treatment significantly increased production of Th 1 type cytokines [IL-2 and interferon (IFN)-${\gamma}$] but not Th 2 type cytokines (IL-4 and IL-10). Therefore, combined treatment with HemoHIM and red ginseng extracts is an effective method to establish powerful immunological synergy in immune cells.

The Mycobacterium avium subsp. Paratuberculosis protein MAP1305 modulates dendritic cell-mediated T cell proliferation through Toll-like receptor-4

  • Lee, Su Jung;Noh, Kyung Tae;Kang, Tae Heung;Han, Hee Dong;Shin, Sung Jae;Soh, Byoung Yul;Park, Jung Hee;Shin, Yong Kyoo;Kim, Han Wool;Yun, Cheol-Heui;Park, Won Sun;Jung, In Duk;Park, Yeong-Min
    • BMB Reports
    • /
    • v.47 no.2
    • /
    • pp.115-120
    • /
    • 2014
  • In this study, we show that Mycobacterium avium subsp. paratuberculosis MAP1305 induces the maturation of bone marrow-derived dendritic cells (BMDCs), a representative antigen presenting cell (APC). MAP1305 protein induces DC maturation and the production of pro-inflammatory cytokines (Interleukin (IL)-6), tumor necrosis factor (TNF)-${\alpha}$, and IL-$1{\beta}$) through Toll like receptor-4 (TLR-4) signaling by directly binding with TLR4. MAP1305 activates the phosphorylation of MAPKs, such as ERK, p38MAPK, and JNK, which is essential for DC maturation. Furthermore, MAP1305-treated DCs transform naive T cells to polarized $CD4^+$ and $CD8^+$ T cells, thus indicating a key role for this protein in the Th1 polarization of the resulting immune response. Taken together, M. avium subsp. paratuberculosis MAP1305 is important for the regulation of innate immune response through DC-mediated proliferation of $CD4^+$ and $CD8^+$ T cells.

Springtime Distribution of Inorganic Nutrients in the Yellow Sea: Its Relation to Water Mass (수괴특성에 따른 춘계 황해의 영양염 분포 특성)

  • Kim, Kyeong-Hong;Lee, Jae-Hak;Shin, Kyung-Soon;Pae, Se-Jin;Yoo, Sin-Jae;Chung, Chang-Soo;Hyun, Jung-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
    • /
    • v.5 no.3
    • /
    • pp.224-232
    • /
    • 2000
  • Inorganic nutrient concentrations in relation to springtime physical parameters of the Yellow Sea were investigated during April 1996. Three major water masses, i.e., the Yellow Sea Warm Current Water (YSWC), Coastal Current Water (CCW) and Changjiang River Diluted Water (CRDW), prevailed in the study area. Water masses were vertically wel1 mixed throughout the study area, and nutrients were supplied adequately from bottom to surface layer. As result of ample nutrients supplied by vertical mixing together with progressed daylight condition, springtime phytoplankton blooms were observed, which was responsible for the depletion of inorganic nutrients in surface water column. Low nutrients concentration in bottom water of the central Yellow Sea (Stn. D9; nitrate: <2 ${\mu}$M, phosphate: <0.3 ${\mu}$) was associated with the entrance of YSWC which is characterized by high temperature and salinity. Influenced by runoff and vertical tidal mixing, CCW with high nutrient concentrations probably associated with China and Korea coastal waters with high nutrients concentration. For the local scale of inorganic nutrient distribution, nutrient transfers from coast to central areas were limited due to restriction imposed by tidal fronts (Stn. D6) and thus affected the horizontal nutrient profiles. Relatively high phytoplankton biomass was observed in the tidal front (Chl-${\alpha}$=12.38 ${\mu}$gL$^{-1}$) during the study period. Overall, the springtime nutrient distribution patterns in the Yellow Sea appeared to be affected by: (1) Large-scale influx of YSWC with low nutrient concentrations and CCW with high nutrient concentrations influenced by Korea and China coastal waters; (2) vertical mixing of water mass and phytoplankton distribution; and (3) local-scale tidal front as well as phytoplankton blooms alongthe tidal front.

  • PDF

Characterization of Interaction Between Porcine Reproductive and Respiratory Syndrome Virus and Porcine Dendritic Cells

  • Park, Jie-Yeun;Kim, Hyun-Soo;Seo, Sang-Heui
    • Journal of Microbiology and Biotechnology
    • /
    • v.18 no.10
    • /
    • pp.1709-1716
    • /
    • 2008
  • The porcine reproductive and respiratory syndrome Virus (PRRSV) is an infectious disease that causes abortions and respiratory disorders in swine. In this study, the interaction between PRRSV and porcine dendritic cells generated from $CD14^{+}$ monocytes in the presence of GM-CSF and IL-4 was examined. As a result, it was shown that immature and mature dendritic cells can be productively infected with PRRSV. When the expression of surface MHC molecules on infected dendritic cells was determined, MHC classes I and II were found to be downregulated when compared with un infected dendritic cells. With the exception of the IL-4 and IFN-$\gamma$ cytokines, the induction of the IL-10, IL-12, and TNF-$\alpha$ cytokines all increased in dendritic cells infected with PRRSV. A mixed lymphocyte reaction showed that peripheral blood mononuclear cells cocultured with PRRSV-infected dendritic cells were less stimulated than peripheral blood mononuclear cells cocultured with dendritic cells treated with PBS, LPS, or UV-inactivated PRRSV. Therefore, these results suggest that PRRSV would appear to modulate the immune stimulatory function of porcine dendritic cells.