• Title/Summary/Keyword: Glycoprotein (G)

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Effects of Transferrin on Enhancing Biological Availability of Iron

  • Park, In-wook;Kim, Yun-Ji;Seong, i-Seung
    • Preventive Nutrition and Food Science
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    • v.3 no.3
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    • pp.248-250
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    • 1998
  • In this study, transferrin which is an iron-carrying glycoprotein in plasma was evaluted for its iron binding capacities(TIBC), iron solubilizing abilities, and enhancing effect of biological availbability of iron. Results of TIBC showed that 1 mg of transferrin could blind 1.28$\mu\textrm{g}$ of iron indicating that one molecule of transferrin can bind about 2 molecules of iron. Also, solubility of iorn (7.5$\mu\textrm{g}$ Fe/ml) was significantly incresed to 96.0% with addition of transferrin (5mg/ml) .When FeCl3(80$\mu\textrm{g}$ Fe/ml) was injected to iron-deficient rats by intestinal segment in situ technique, 18.4% of injected iron was absorbed wherease 48.49 and 48.76% of injected iron was absorbed with addition of 10 and 20 mg transferrin/ml , respectively.

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Effect of Organic Farming on Spore Diversity of Arbuscular Mycorrhizal Fungi and Glomalin in Soil

  • Lee, Ji-Eun;Eom, Ahn-Heum
    • Mycobiology
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    • v.37 no.4
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    • pp.272-276
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    • 2009
  • In this study, eight soil samples were collected from organic and conventional farms in a central area of South Korea. Spore communities of arbuscular mycorrhizal fungi (AMF) and glomalin, a glycoprotein produced by AMF, were analyzed. Spores of Glomus clarum, G. etunicatum, G. mosseae, G. sp., Acaulospora longula, A. spinosa, Gigaspora margarita, and Paraglomus occultum were identified at the study sites, based on morphological and molecular characteristics. While Acaulospora longula was the most dominant species in soils at organic farms, Paraglomus occultum was the most dominant species in soils at conventional farms. Species diversity and species number in AMF communities found in soils from organic farms were significantly higher than in soils from conventional farms. Glomalin was also extracted from soil samples collected at organic and conventional farms and was analyzed using both Bradford and enzyme-linked immunosorbent assays. The glomalin content in soils from organic farms was significantly higher than in soils from conventional farms. These results indicate that agricultural practices significantly affect AMF abundance and community structure.

Isolation of Anticarcinogenic Isoflavone-conjugated Glycoproteins from a Submerged Liquid Culture of Agaricus blazei Mycelia by the Autolysis Process (신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리)

  • Kim, So Young;Kim, Young Suk;Jang, Joung Soon;Kim, Boh Hyun;Rakib, Abdur Md.;Kim, Gon Sup;Kim, Jeong Ok;Ha, Yeong Lae
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1316-1324
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    • 2014
  • Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at $53^{\circ}C$ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with $^1H$-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.

Autophagy-Dependent Survival of Mutant B-Raf Melanoma Cells Selected for Resistance to Apoptosis Induced by Inhibitors against Oncogenic B-Raf

  • Ahn, Jun-Ho;Lee, Michael
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.114-120
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    • 2013
  • Most patients with mutant B-Raf melanomas respond to inhibitors of oncogenic B-Raf but resistance eventually emerges. To better understand the mechanisms that determine the long-term responses of mutant B-Raf melanoma cells to B-Raf inhibitor, we used chronic selection to establish B-Raf (V600E) melanoma clones with acquired resistance to the new oncogenic B-Raf inhibitor UI-152. Whereas the parental A375P cells were highly sensitive to UI-152 ($IC_{50}$ < $0.5{\mu}M$), the resistant sub-line (A375P/Mdr) displayed strong resistance to UI-152 ($IC_{50}$ < $20{\mu}M$). Immunofluorescence analysis indicated the absence of an increase in the levels of P-glycoprotein multidrug resistance (MDR) transporter in A375P/Mdr cells, suggesting that resistance was not attributable to P-glycoprotein overexpression. In UI-152-sensitive A375P cells, the anti-proliferative activity of UI-152 appeared to be due to cell-cycle arrest at $G_0/G_1$ with the induction of apoptosis. However, we found that A375P/Mdr cells were resistant to the apoptosis induced by UI-152. Interestingly, UI-152 preferentially induced autophagy in A375P/Mdr cells but not in A375P cells, as determined by GFP-LC3 puncta/cell counts. Further, autophagy inhibition with 3-methyladenine (3-MA) partially augmented growth inhibition of A375P/Mdr cells by UI-152, which implies that a high level of autophagy may protect UI-152-treated cells from undergoing growth inhibition. Together, our data implicate high rates of autophagy as a key mechanism of acquired resistance to the oncogenic B-Raf inhibitor, in support of clinical studies in which combination therapy with autophagy targeted drugs is being designed to overcome resistance.

Inhibition of Human $CD8^+$ Cytotoxic T Lymphocyte (CTL) -mediated Cytotoxicity in Porcine Fetal Fibroblast Cells by Overexpression of Human Cytomegalovirus Glycoprotein Unique Short (US) 2 Gene

  • Park, K-W.;Yoo, J.Y.;Choi, K.M.;Yang, B.S.;Im, G.S.;Seol, J.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.20-25
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    • 2009
  • Xenotransplantation of pig organs into humans is a potential solution for the shortage of donor organs for transplantation. However, multiple immune barriers preclude its clinical application. In particular, the initial type of rejection in xenotransplantation is an acute cellular rejection by host $CD8^+$ cytotoxic T lymphocyte (CTL) cells that react to donor major histocompatibility complex (MHC) class I. The human cytomegalovirus (HCMV) glycoprotein Unique Short (US) 2 specifically targets MHC class I heavy chains to relocate them from the endoplasmic reticulum (ER) membrane to the cytosol, where they are degraded by the proteasome. In this study we transfected the US2 gene into minipig fetal fibroblasts and established four US2 clonal cell lines. The integration of US2 into transgenic fetal cells was confirmed using PCR and Southern blot assay. The reduction of Swine Leukocyte Antigen (SLA)-I by US2 was also detected using Flow cytometry assay (FACS). The FACS analysis of the US2 clonal cell lines demonstrated a substantial reduction in SLA-I surface expression. The level (44% to 76%) of SLA-I expression in US2 clonal cell lines was decreased relative to the control. In cytotoxicity assay the rate of $CD8^+$ T cell-mediated cytotoxicity was significantly reduced to 23.8${\pm}$15.1% compared to the control (59.8${\pm}$8.4%, p<0.05). In conclusion, US2 can directly protect against $CD8^+$-mediated cell lysis. These results indicate that the expression of US2 in pig cells may provide a new approach to overcome the CTL-mediated immune rejection in xenotransplantation.

Cloning of the non-virion (NV) of a Korean Isolate of Infectious Hematopoietic Necrosis and Identification of the Role of the NV in IHNV Replication (한국에서 분리된 전염성 조혈괴저 바이러스의 non-virion (NV) 단백질의 유전자 클로닝 및 바이러스 증식에서의 역할)

  • 문창훈;조화자;윤원준;박정재;박정민;김현주;도정완;이주양;임채렬
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.103-108
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    • 2000
  • We have cloned and analyzed cDNA coding for non-virion (NV) protein of the m V - P R T The NV gene contained 336 bp open readmg frame and encoded a protein of 11 1 amino acids with a molecular weight of 13.2 kDa. The deduced amino acid sequence of NV of IHNVPRT was found to be 90-95% identical to those of foreign isolates of IHNV. These results indicate that NV gene of the MNV is highly conserved among &ifferent strains of THNV Northern blot analyses revealed that the levels of NV gene expression were strongly elevated after 20 h post-infection. In order to identify the role of NV in the replication of MNV in fish cells, IHNVinfected cells were treated with antisense oligonucleotides. While IHNV-PRT exposed to glycoprotein (G) antisense oligonucleotide showed severely reduced growth, the growth of virus exposed to NV antisense oligonucleotide was not affected by NV antisense oligonucleotide, which suggests that NV is not essential for replication of IHNV in fish cells.

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Monitoring of VHS and RSIVD in cultured Paralichthys olivaceus of Jeju in 2014 (2014년 제주도 양식 넙치, Paralichthys olivaceus에 대한 VHS 및 RSIVD 모니터링)

  • PARK, Hyun-Kyung;KIM, Seung-Min;LEE, Da-Won;JUN, Lyu-Jin;JEONG, Joon-Bum
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.3
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    • pp.879-889
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    • 2015
  • The outbreak of viral diseases caused by viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) have been reported in cultured olive flounder, Paralichthys olivaceus. VHSV has been a serious viral disease that infects the olive flounders in South Korea. Clinical signs of VHSV infection are skin darkening, abdominal distension and haemorrhages. Outbreaks of fish iridoviral disease was first reported from red seabream, Pagrus major farms in Japan. Recently, iridovirus infection have occurred frequently from olive flounder farms in South Korea. In this study, disease surveillance was performed to monitor the prevalence of VHSV and RSIV in olive flounder in 2014. The samples were collected from 60 different olive flounder farms in Jeju from April, May, September, November and December in 2014. RT-PCR (VHSV) or PCR (RSIV) results showed that VHSV were detected in 5 farms, but RSIV has not been detected in any farms. The migration of olive flounder was restricted for the quarantine in 5 farms of VHS outbreak. The nucleocapsid protein (N) gene and glycoprotein (G) gene sequences of the 5 Korean VHSV isolates were successfully amplified and sequenced. Phylogenetic analysis was performed using the VHSV sequences reported here together comparison with the nucleotide sequences available from the GenBank database. Phylogenetic analysis indicated that most of Korea VHSV belong to the genotype IVa and closely related to the strains from Japan and China.

Detection of viral hemorrhagic septicemia virus (VHSV) in wild marine fishes in the coastal region of Korea (우리 나라 연근해 자연산 해수 어종에서의 Viral Hemorrhagic Septicemia Virus (VHSV)의 검출)

  • Kim, Su-Mi;Park, Su-Il
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.1-10
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    • 2004
  • In order to analyse the detection of viral hemorrhagic septicemia virus (VHSV) in marine environment surrounding coastal region of Eastern and Southern sea of Korea, the pools of each organ sample of three fish were taken for virus assay from February to May in 2003. The samples comprised 42, taken from 9 species of marine fishes. The VHSV was detected from chub mackerel Scomber japonicus and striped mullet Mugil cephalus in epithelioma papulosum cyprini (EPC) cells. The identity of the virus was confirmed a reverse transcriptase-polymerase chain reaction (RT-PCR). VHSV has previously been reported from chub mackerel, but not from striped mullet. The new isolates was classified as a member of genogroup I (American type) of VHSV and was closely related to the VHSV KVHS'01-l based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene.

Isolation and Characterization of Immunomodulatory Glycoprotein from the Root of Panax ginseng

  • Shin, Han-Jae;Park, Kyeong-Mee;Kim, Young-Sook;Nam, Ki-Yeul;Lee, You-Hui;Park, Jong-Dae
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.128-133
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    • 2000
  • A high molecular (more than 10 kDa) fraction, showing mitogenic and comitogenic activities in spleen cells of mouse, was isolated from water extract of ginseng. The crude protein substance prepared by 80% (NH$_4$)$_2$SO$_4$ precipitation from this fraction was purified and isolated by DEAE Sepharose column chromatography. Among the fractions eluted, it was found that four kinds of fractions eluted with 0 to 1 M NaCl gradient were glycoproteins, which induced proliferation of spleen cells and increased NO production in macrophages. Among them, F-2 fraction, which contained 35.9% protein,49.4% neutral sugar and 12.5% uronic acid, was found to show mitogenic activity as strong as that of LPS (lipopolysaccharide) at a concentration of 100 $\mu\textrm{g}$/ml and to remarkably stimulate NO production by murine macrophages at a concentration of 500 $\mu\textrm{g}$/ml. When F-2 is deproteinized, the mitogenic activity of F-2 was decreased significantly to 70.9% as compared with that of F-2. This results suggests that the protein moiety of F-2 may play an important role in immunomodulating activity of glycoprotein from the root of Panax ginseng.

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Electrophoretic analysis of the major proteins of ruminant erythrocyte membrane: Their relation to slow erythrocyte sedimentation rate (반추동물 적혈구막 단백의 전기영동법에 의한 분석 -낮은 적혈구침강속도와의 관계-)

  • Lee, Bang-whan;Bahk, Young-woo
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.445-455
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    • 1989
  • The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate(ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was $2.85{\pm}0.28$ in human, $3.60{\pm}0.41$ in Korean cattle, $3.71{\pm}0.36$ in Holstein, $4.13{\pm}0.83$ in Korean native goat and $3.94{\pm}0.56mg/ml$ in sheep, showing higher in ruminant animals than in human(p<0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band-Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycogrotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep. It was particularly noticeable that PAS-B, a fraction of glycoprotein, present in ruminants except sheep, was better digested by proteinases than by glycosidases, showing remarkable increase(p<0.01) of the ESR in accord with complete digestion(disappearance) of the PAS-B band by pronase, trypsin or chymotrypsin treatment of erythrocytes. In sheep, there was almost no any response to the various enzymes in general protein and glycoprotein profiles of the erythrocyte membranes except PAS-G, which was markedly decreased by pronase treatment of the erythrocytes. Nevertheless, the ESRs were accelerated in erythrocytes treated with pronase, trypsin, chymotrypsin and neuraminidase. Erythrocyte osmotic fragility was increased in erythrocytes treated with only pronase among five enzymes in all the human and ruminant animals used in this study.

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