• Title/Summary/Keyword: Glycoprotein

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Analysis of antigenicity of viral hemorrhagic septicemia virus (VHSV) glycoprotein from cultured olive flounder Paralichthys olivaceus (양식 넙치, Paralichthys olivaceus에서 분리한 viral hemorrhagic septicemia virus (VHSV)의 항원성 분석)

  • Kim, Su-Mi;Jee, Bo-Young;Cho, Mi-Young;Won, Kyoung-Mi;Kim, Jin-Woo;Park, Soo-Il
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.75-84
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    • 2011
  • The amino acid sequence of glycoprotein of Korean VHSV isolate (KR'01-1) was analyzed using the DNAStar Protean system. Based on the flexibility, hydrophilicity, antigenic index and surface probability, three regions (Gp1, Gp2 and Gp3) were selected as potential antigenic determinants. Three oligopeptides containing the amino acid sequences of the three regions were synthesized and polyclonal antibodies were raised against them. The activities of the antibodies were analyzed by Western blotting and virus neutralization test. The results showed that antibodies raised against oligopeptides Gp1 and Gp2 neutralized the infectivity of VHSV, suggesting that they can be possible candidates for subunit vaccines against VHS diseases in olive flounder.

Cloning of cDNA Encoding PAS-4 Glycoprotein, an Integral Glycoprotein of Bovine Mammary Epithelial Cell Membrane

  • Hwangbo, Sik;Lee, Soo-Won;Kanno, Chouemon
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.576-584
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    • 2002
  • Bovine PAS-4 is an integral membrane glycoprotein expressed in mammary epithelial cells. Complementary DNA (cDNA) cloning of PAS-4 was performed by reverse-transcriptase polymerase chain reaction (RT-PCR) with oligonucleotide probes based on it's amino terminal and internal tryptic-peptides. The cloned PAS-4 cDNA was 1,852 nucleotides (nt) long and its open reading frame (ORF) was encoded 1,413 base long. The deduced amino acid sequence indicated that PAS-4 consisted of 471 amino acid residues with molecular weight of 52,796, bearing 8 potential N-glycosylation sites and 9 cysteine residues. Partial bovine CD36 cDNA from liver also was sequenced and the homology of both nucleotide sequence was 94%. Most of the identical amino acid residues were in the luminal/extracellular domains. Contrary to PAS-4, bovine liver CD36 displays 6 potential N-glycosylation sites, which were located, except for those at positions 101 and 171, at same positions as PAS-4 cDNA. Cysteine residues of PAS-4 and CD36 were same at position and in numbers. Northern blot analysis showed that PAS-4 was widely expressed, although its mRNA steady-state levels vary considerably among the analyzed cell types. PAS-4 possessed hydrophobic amino acid segments near the amino- and carboxyl-termini. Two short cytoplasmic tails of the amino- and carboxyl-terminal ends constituted of a 5-7 and 8-11 amino acid residues, respectively.

A Systematic Review on Potential Drug Interactions of Korean Ginseng Based upon Cytochrome P450 and P-Glycoprotein (고려인삼의 Cytochrome P450 및 P-Glycoprotein 매개 약물 상호작용에 미치는 영향)

  • Nam, Ki Yeul;Yang, Byung Wook;Shin, Wang Soo;Park, Jong Dae
    • Korean Journal of Pharmacognosy
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    • v.49 no.2
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    • pp.85-102
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    • 2018
  • A drug interaction is a situation in which a substance affects the activity of a drug, synergistically or antagonistically, when both are administered together. It has been shown that orally taken ginsenosides are deglycosylated by intestinal bacteria to give ginsenosides metabolites, which has been considered to be genuine pharmacological constituents and to exhibit drug interactions. Animal experimental results demonstrated that ginsenoside metabolites play an important role in the inhibitory or inductive action of both CYPs (cytochrome p450) and P-gp (p-glycoprotein), thereby can be applied as metabolic modulator to drug interactions. Very few are known on the possibility of drug interaction if taken the recommended dose of ginseng, but it has been found to act as CYPs inductor and P-gp inhibitor in any clinical trial, suggesting the risk that side effects will occur. It has been recently reported that interactions might also exist between ginseng and drugs such as warfarin, phenelzine, imatinib and raltegravir. Moreover, medicinal plants are increasingly being taken in a manner more often associated with prescription medicines. Therefore, considering the extensive applications of ginseng for safety, the aim of this review is to present a comprehensive overview of ginseng and drug interactions based upon pharmacodynamic and pharmacokinetic evidences.

Gene Transfer into Chicken Embryos using Defective Retroviral Vectors Packaged with Vesicular Stomatitis Virus G Glycoprotein Envelopes (Vesicular Stomatitis Virus G Glycoprotein Envelope으로 포장된 Defective Retroviral Vector를 이용한 닭의 배로의 유전자 전이)

  • 권모선;임은정;허영태;이훈택;이영만;김태완
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.171-180
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    • 2001
  • Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10$^{8}$ CFU/ $m\ell$ on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100$\times$) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.

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Analysis of Non-segregated S-allele Strain by Single-Locus Hypothesis in Self-incompatible Brassica campestris (자가불화합성 Brassica campestris에 있어서 단일유전자좌가설에 의해 분리되지 않는 S-유전자 계통의 분석)

  • 노일섭
    • Journal of Plant Biology
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    • v.36 no.2
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    • pp.127-132
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    • 1993
  • Self-incompatibility in Brassica campestris is controlled by multi-allele system in a single genetic locus, the S locus, and it is elucidated that S-glycoproteins are S gene products. In this experiments, we examined the genetic mode(pollen tube behavior and segregation of S-glycoprotein), characteristic of S-glycoproteins and DNA constitution within nuclear genome on S gene family that unexplained by single locus model, and investigated the segregation pattern of S-glycoproteins in bred F1 generation. By diallel cross among the 15 plants within one family the existence of three types of homozygotes and three types of heterozygotes were observed, and segregation of S-allele could not explained by single locus model. From the results of IEF-immunoblot analysis for non-segregated individual plant, the segregation pattern of S specific bands was corresponded with results of diallel cross except with one case(SaSa genotype). The molecular weight of 6 different S-genotype varied in near by 50 kD, and each genotype expressed with 2 or 3 bands. Specific bands in SaSa, SbSb, ScSc has almost similar molecular weight between them. Southern analysis of genomic DNA probed with S-glycoprotein cDNA for 6 different genotypes revealed that there are clear difference in polymorphism, multiple bands of hybridization, when restriction enzymes of EcoR I were used. It could be assumed that there are several sequences related to the S-glycoprotein structural genes within their nuclear genome. Therefore, we suggested the possibilities that S-allele system could be controlled by multi-locus, that dominance-recessive interactions could be explained by modifier gene or supressor gene based on the results of abnormal segregation of S-glycoprotein in bred F1. The F2 analyses are progressing in now.

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Anti-Metastatic Activity of Glycoprotein Fractionated from Acanthopanax senticosus, Involvement of NK-cell and Macrophage Activation

  • Ha, Eun-Suk;Hwang, Soo-Hyun;Shin, Kwang-Soon;Yu, Kwang-Won;Lee, Keyong-Ho;Choi, Joo-Sun;Park, Woo-Mun;Yoon, Taek-Joon
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.217-224
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    • 2004
  • Previously, we reported that water-extracted Acanthopanax senticasus exhibited anti-meta-static activity by stimulating the immune system. In this study, we fractionated glycoproteins (EN-SP) from the soluble protein layer (GF-AS) of A. senticasus and determined their basic chemical properties. We also investigated the anti-tumor and immunostimulating activities of the fractionated glycoprotein, EN-SP. We found that intravenous (i.v.) administration of GF-AS dramatically inhibited metastasis of colon26-M3.1 carcinoma cells to the lung in a dose-dependent manner. In vitro analysis showed GF-AS to enhance the proliferation of splenocytes. GF-AS also stimulated peritoneal macrophage, which was followed by the production of various cytokines such as IL-1$\beta$, TNF-$\alpha$, IL-12 and IFN-${\gamma}$. Furthermore, the production of these cytokines was partially blocked when peritoneal macrophage was cultured with the polyclonal antibodies against GF-AS. The depletion of NK cells by rabbit anti-asialo GM1 serum partly abolished the inhibitory effect of GF-AS on lung metastasis of colon26-M3.1 cells. Using gel filtration, EN-SP, an active glycoprotein fraction, is isolated from GF-AS. While both GF-AS and EN-SP stimulated the proliferatation of splenocytes of normal mice, EN-SP showed higher anti-metastatic activity and more potently stimulated the proliferation of splenocytes compared to GF-AS. These results suggest the use of EN-SP, the fractionated glycoprotein from A. senticasus, can be used as a therapeutical reagent to prevent or inhibit tumor metastasis.

Effects of Nimodipine on the Pharmacokinetics of Warfarin in Rats: A Possible Role of P-glycoprotein and CYP3A4 Inhibition by Nimodipine (와파린의 약물동태학에 니모디핀의 영향)

  • Moon, Hong Seop;Lee, Chong Ki;Burm, Jin Pil
    • Korean Journal of Clinical Pharmacy
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    • v.23 no.3
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    • pp.206-212
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    • 2013
  • Purpose: The aim of this study was to investigate the effect of nimodipine on the pharmacokinetics of warfarin after oral and intravenous administration of warfarin in rats. Methods: Warfarin was administered orally (0.2 mg/kg) or intravenously (0.05 mg/kg) without or with oral administration of nimodipine (0.5 or 2 mg/kg) in rats. The effect of nimodipine on the P-glycoprotein as well as cytochrome P450 (CYP) 3A4 activity was also evaluated. Results: Nimodipine inhibited CYP3A4 enzyme activity with 50% inhibition concentration ($IC_{50}$) of $10.2{\mu}M$. Compared to those animals in the oral control group (warfarin without nimodipine), the area under the plasma concentration-time curve (AUC) of warfarin was significantly greater (0.5 mg/kg, P<0.05; 2 mg/kg, P<0.01) by 31.3-57.6%, and the peak plasma concentration ($C_{max}$) was significantly higher (2 mg/kg, P<0.05) by 29.4% after oral administration of warfarin with nimodipine, respectively. Consequently, the relative bioavailability of warfarin increased by 1.31- to 1.58-fold and the absolute bioavailability of warfarin with nimodipine was significantly greater by 64.1-76.9% compared to that in the control group (48.7%). In contrast, nimodipine had no effect on any pharmacokinetic parameters of warfarin given intravenously. Conclusion: Therefore, the enhanced oral bioavailability of warfarin may be due to inhibition of CYP 3A4-mediated metabolism rather than P-glycoprotein-mediated efflux by nimodipine.

Expression and Characterization of Human Immunodeficiency Virus-1 Oligomerized gp140 Protein in Mammalian Cells (포유동물 세포에서 Human Immunodeficiency Virus-1의 Oligomeric gp140 단백의 발현 및 특성)

  • Kim, Eun-Ok;Kim, Eun;Kim, Hyun-Soo;Shin, Kwang-Soon;Kim, Chul-Joong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.55-64
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    • 2002
  • The envelope glycoprotein of HIV-1 forms an oligomeric complex resulting in playing a role to induce neutralizing antibody and cell-mediate immune responses. The oligomer exists as a trimer of gp120-gp41 heterodimer which mediates HIV-1 attachment and fusion. We made a cDNA clone of gp140 consisting of gp120 and ectodomain of gp41 from the primary African isolate. To express the oligomeric gp140 in mammalian cells, we adopted the Semliki Forest virus (SFV) based expression system. The oligomeric gp140 in the secretory form was expressed and purified from the cell culture supernatant and characterized. The antibody inducing activity of the purified gp140 was also examined in mice inoculation.

Establishment of multiplex RT-PCR for differentiation between rabies virus with and that without mutation at position 333 of glycoprotein

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Yoo, Jae-Young
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.22.1-22.9
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    • 2020
  • Rabid raccoon dogs (Nyctereutes procyonoides koreensis) have been responsible for animal rabies in South Korea since the 1990s. A recombinant rabies vaccine strain, designated as ERAGS, was constructed for use as a bait vaccine. Therefore, new means of differentiating ERAGS from other rabies virus (RABV) strains will be required in biological manufacturing and diagnostic service centers. In this study, we designed two specific primer sets for differentiation between ERAGS and other RABVs based on mutation in the RABV glycoprotein gene. Polymerase chain reaction analysis of the glycoprotein gene revealed two DNA bands of 383 bp and 583 bp in the ERAGS strain but a single DNA band of 383 bp in the field strains. The detection limits of multiplex reverse transcription polymerase chain reaction (RT-PCR) were 80 and 8 FAID50/reaction for the ERAGS and Evelyn-Rokitnicki-Abelseth strains, respectively. No cross-reactions were detected in the non-RABV reference viruses, including canine distemper virus, parvovirus, canine adenovirus type 1 and 2, and parainfluenza virus. The results of multiplex RT-PCR were 100% consistent with those of the fluorescent antibody test. Therefore, one-step multiplex RT-PCR is likely useful for differentiation between RABVs with and those without mutation at position 333 of the RABV glycoprotein gene.

Protein Patterns of Ovary and Hemolvmph in Gerris paludum

  • 황명실;최원철
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.337-350
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    • 1989
  • In this study, we compared patterns of hemolvmph with ovarian proteins during the yolk formation of Gewis paludum by using SDS/polyacrylamide gel electrophoresis and he dimen-tlonal electrophoresis. In addition, we examined patterns of glycoprotein which were composed of yolk substances. The results were as follows: The protein patterns in ovary of the 5th instar without eggs were similar to those of adult after ovulation. Protein amounts of the ovaw without developing eggs were less than those of the ovaw containing oocvtes or matured eggs at the molecular weights range from 66, 000 to 110.000 daltons. No glvcoproteins were observed in ovaw without eggs. But the glvcoproteins were gradually increased according to development of eggs in the 5th instar. After the ovulated ovaw of adult, no glvcoprotein bands urere occurred as the bands of the ovary without eggs in the 5th instar. Also, amounts of hemolvmph protein between 66, 000 and 110, 000 in molecular weight were increased during yolk formation of the 5th instal. The results suggest that ovarian protein substances may originate from hemolpnph. In the 5th instar, the amounts of glycoprotein of developing eggs was gradually increased in the hemolymph. The band of M.W. 29, 500 was occurred in the hemolpnph of the 5th instar and adult without eggs. This protein mal be precursor of glvcoprotein in the high molecular weight area in the ovary of more developed eggs. The number of spots and the amounts of protein in ovary without eggs were less than those of ovary containing eggs by two dimensional elec-trophoresis. The protein bands between 45, 000 and 110, 000 almost appeared in acidic field of he dimensional gel. Especially, the band, M.W. 109, 000, uras separated 3 spots, a1, a2, and as. The band, M.W. 102, 000, has spots which designated b1, b2, b3, and b4. Besides, proteins below M.W. 24, 000 were occurred less spots in the basic field than those of acidic field. The mechanism of intracellular organs (= cell organells) uras related to the yolk protein synthesis of oocyte in the process of yolk protein formation of derris pcfudum.

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