• Title/Summary/Keyword: Glycoproteins

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Expression and Characterization of the Human Immunodeficiency Virus Type 1 Mutant Envelope Glycoproteins in Mammalian Cells (진핵세포에서 HSV-1 Envelope 변이 단백질의 발현 및 발현 단백질의 특성 연구)

  • Ryu, Ji-Yoon;Park, Jin-Seu
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.183-193
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    • 1999
  • Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

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Study on the Expectorant Activity of Ethanol Extracts from Some Crude Drugs Containing Saponins (Saponin을 함유한 생약추출물의 거담작용에 관한 연구)

  • Kim, Sook-Young;Moon, Ja-Yeong;Lee, Dong-Wook;Park, Ki-Hyun
    • Korean Journal of Pharmacognosy
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    • v.19 no.2
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    • pp.133-140
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    • 1988
  • The effect of ethanol extracts of Gilkyung(Platicodi Radix, Platicodongrandiflorum A, DC), Onji(Polygalae Radix, Polygala tennuifolia Willdenow) and Deoduk(Codonopsis lanceolate Radix, Codonopsis lanceolata) on expectorant activity of rat trachea was investigated. Following treatment of 50% ethanol extract of these medicinal plants (25 mg/rat), the content of neurotransmitters such as acetylcholine and histamine in tissue was significantly increased. The secretions of acid glycoproteins and the artificially injected phenol red were also increased. However, there was no significant difference except Onji From the histological study through periodic acid Schiff and alcian blue stain, the thickness of inner membrane of acinar glands and the stained glycoproteins on surface of epithelium and on the glands were observed in all the rats trachea treated with extract of medicinal plants. In vitro, the viscosity of mucin solution was slightly decreased with an addition of the extracts. Onji showed the most effective expectorant activity among them at the identical conditions. The mechanism of expectorant activity of these medicinal plants seems to be due to stimulation of secretion and changes of rheological properties of mucus.

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Specificity in the Inhibition of Mucin Release from Airway Goblet Cells by Polycationic Peptides (호흡기 배상세포에서 폴리양이은성 펩티드에 의해 야기되는 뮤신유리 억제 현상의 특이성 규명 Specificity in the Inhibition of Mucin Release from Airway Goblet Cells by Polycationic Peptides)

  • 이충재
    • Biomolecules & Therapeutics
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    • v.9 no.3
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    • pp.218-223
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    • 2001
  • In the present study, we intended to investigate whether polycationic peptides including poly-L-lysine (PLL) and poly-L-arginine (PLA) specifically inhibit the mucin release and do not affect significantly the release of the other releasable glycoproteins with less molecular weight than mucin's from cultured airway goblet cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hr and chased for 30 min in the presence of varying concentrations of either poly-L-arginine (PLA) or poly-L-lysine (PLL) to assess the effects on 3H-mucin release and on the total elution profile of the treated culture medium. The results were as follows : (1) PLL 78,000, PLL 9,600 and PLA 8,900 inhibited mucin release in a dose-dependent manner; (2) These polycationic peptides did not inhibit the release of the other releasable glycoproteins with less molecular weights than mucin's. We conclude that these polycationic peptides 'specifically'inhibit mucin release from airway goblet cells. This finding suggests that these polycationic peptides might be used as a specific airway mucin-regulating agent.

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A study on several new glycoproteins isolated from crude soybean hemagglutinin (Soybean Hemagglutinin의 정제중(精製中) 분리(分離)되는 Glycoprotein에 관(關)한 연구(硏究))

  • Kim, S.I.;Lee, C.Y.
    • Applied Biological Chemistry
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    • v.12
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    • pp.1-5
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    • 1969
  • 1. The purified soybean hemagglutinin isolated from Korean soybean through the calcium phosphate column chromatography was found to contain mannose (4.9%) and glucosamine (1.1%) added to the protein portion. 2. The fractions I,II,III, the impurities accompanied by hemagglutinin during the final purification were proved to be new plant glycoproteins that were seemingly paper electrophoretically homogeneous and had the same mobility. 3. The carbohydrate and nitrogen contents of I,II,III were 4.5%, 1.13%, 1.1% in mannose, 0.5%, 1.2%, 1.22% in glucosamine and 12.8%, 15.5%, 13.9% in nitrogen respectively.

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Modification of carbohydrate compositions of 31/36 kDa proteins of plerocercoids (sparganum) of Spirometra mansoni grown in different intermediate hosts

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.77-79
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    • 2004
  • We purified specific 31/36 kDa antigenic molecules from sparganum in different intermediate hosts (snakes and mice) and analyzed their monosaccharide compositions. Compositional analysis showed that glucose and man nose concentrations were 2-3 fold higher in the 31/36 kDa molecule purified from snakes than those from mice. This result implies that antigenic glycoproteins of sparganum from snakes might be modified in mammalian sparganosis with respect to their carbohydrate composition.

Immunohistochemical detection of infectious hematopoietic necrosis virus antigens in cell cultures (배양세포에서 전염성조혈장기괴사증 바이러스항원의 면역조직화학적 검출)

  • 문운경;이민권;진영배;김순복
    • Korean Journal of Veterinary Service
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    • v.25 no.3
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    • pp.295-297
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    • 2002
  • This experiment was done to set up the immunohistochemical detection method for infectious hematopoietic necrosis virus(IHNV) antigens in the monolayers of CHSE-214 cell cultures inoculated with IHNV. Specific identification of IHNV antigens was detected in the cytoplasms of infected cells by the use of monoclonal antibodies to glycoproteins. The specific positive signal was observed as a distinct red color. The result showed that streptavidin alkaline phosphatase immunohistochemistry specifically identified IHNV antigens in infected cultured cells.

Total synthesis of 1,4-Dideoxy-1,4-Imino-D-Arabinitol(DABl)

  • Kim, In-Su;Jung, Young-Hoon
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.178.2-178.2
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    • 2003
  • Naturally occurring sugar mimics with a nitrogen in the ring are classified into five structural classes: polyhydroxylated pyrrolidines, piperidines, indolizidines. pyrrolizidine, and nortropanes. Glycosidase are involved in a wide range of important biological processes, such as intestinal digestion, post-translational processing of glycoproteins and the lysosomal catabolism of glycoconjugate. The realization that alkaloidal sugar mimics might have enormous therapeutic potential in many diseases such as viral infection, cancer and diabetes has led to increasing interest and demand for these compounds. (omitted)

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Cytochemical and Biochemical Characteristics of Cellular Adhesion in Amoeba proteus (Amoeba proteus의 표면흡착에 관한 세포화학 및 생화학적 특성)

  • 안태인;곽인희
    • The Korean Journal of Zoology
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    • v.29 no.3
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    • pp.171-180
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    • 1986
  • The effects of proteases, neuraminidase and EDTA on adhesion of amoebae on the substratum, ultrastructure and biochemical composition of the cell surface were studied by concanavalin A (con A) cytochemistry and SDS PAGE. By con A cytochemistry the glycocalyx of the plasmalemma was easily subdivided into outer filamentous (F) layer and the inner amorphous (A) layer. On treatment with neuraminidase, amoebae attached to the substratum and spreaded better than untreated cells exposing the more con A binding sites in A- and F-layer. When the cells were treated with trypsin or proteinase K, cells stayed unattached for 12 and 48 hr, respectively. Con A binding sites of A layer and all of those glycoproteins were removed by proteinase K. On the other hand, trypsin damaged all of the con A binding sites in both A- and F-layer without significant change in PAS-stained profile of the plasmalemma. Some of the mucopolysaccharides of the cell surface were released by these enzymes and EDTA. When the cells were incubated with monovalent con A they did not attch on the substratum and cytolysed. From these results adhesion of amoebae on the substratum appears to be mediated by the interaction of the glycoproteins and mucopolysaccharides of the A layer.

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Glycoantigen Biosyntheses of Human Hepatoma and Colon Cancer Cells are Dependent on Different N-Acetylglucosaminyltransferase-III and -V Activities

  • Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.891-900
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    • 2004
  • UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.