• Title/Summary/Keyword: Glycoproteins

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Characterization of the $\alpha$-mannosidase Gene Family in Filamentous Fungi: N-glycan Remodelling for the Development of Eukaryotic Expression Systems

  • Eades, C.Joshua;Hintz, William E.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.227-233
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    • 2000
  • Although filamentous fungi are used extensively for protein expression, their use for the production of heterologous glycoproteins is constrained by the types of N-glycan structures produced by filamentous fungi as compared to those naturally found on the glycoproteins. Attempts are underway to engineer the N-glycan synthetic pathways in filamentous fungi in order to produce fungal expression strains which can produce heterologous glycoproteins carrying specific N-glycan structures. To fully realize this goal, a detailed understanding of the genetic components of this pathway in filamentous fungi is required. In this review, we discuss the characterization of the $\alpha$-mannosidase gene family in filamentous fungi and its implications for the elucidation of the N-glycan synthetic pathway.

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Differentiation of Glycan Diversity with Serial Affinity Column Set (SACS)

  • Shin, Jihoon;Cho, Wonryeon
    • Mass Spectrometry Letters
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    • v.7 no.3
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    • pp.74-78
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    • 2016
  • Targeted glycoproteomics is an effective way to discover disease-associated glycoproteins in proteomics and serial affinity chromatography (SAC) using lectin and glycan-targeting antibodies shows glycan diversity on the captured glycoproteins. This study suggests a way to determine glycan heterogeneity and structural analysis on the post-translationally modified proteins through serial affinity column set (SACS) using four Lycopersicon esculentum lectin (LEL) columns. The great advantage of this method is that it differentiates between glycoproteins on the basis of their binding affinity. Through this study, some proteins were identified to have glycoforms with different affinity on a single glycoprotein. It will be particularly useful in determining biomarkers in which the disease-specific feature is a unique glycan, or a group of glycans.

Alterations in Cellular and Plasma Membrane Glycopreteins in Chicken Myogenesis in Vitro (鷄胚의 細胞胞分化에 있어서 細胞 및 原形質膜 糖蛋白質의 變化)

  • Kang, Man-Sik;Song, Woo-Keun;Ho. Woo Nam;Chung, Chang-Yong
    • The Korean Journal of Zoology
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    • v.28 no.3
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    • pp.125-136
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    • 1985
  • The present study has been undertaken to elucidate whether there are developmentally regulated proteins in chick myogenesis by observing cellular and plasma membrane glycoproteins by use of labelled-Con A staining. At least three classes and five classes of glycoproteins can be identified in terms of their occurrence and their change in levels, respectively. As to the former, there are glycoproteins which are specific to cellular fraction are eight, those which are specific to plasma membrane are four, and those which occur in both fraction are nine. Likewise, as to the latter, there are glycoproteins which are invariant, those which increase in level, those which decrease, those which increase and subsequently decrease, and those which decrease and subsequently increase. The present experiment reveals that the level of fibronectin does decrease after the fusion and that the apparent conflicting results so far reported may be due to the difference in time point setting during fusion and myotube stages. It is thought to be a general tendency that glycoproteins of high-molecular-weight decrease and those of low-molecular-weight increase in relative levels as the fusion proceeds. These results are suggestive of the fact that a major structural reorganization of glycoproteins does occur in association with myoblast fusion.

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Sialylated oligosaccharide analyses using high-performance liquid chromatography with a fluorescence detector

  • Cho, Due-Hyeon;Ryu, Chang-Soo;Park, Jee-Hun;Kim, Ha-Hyung
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.316.1-316.1
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    • 2003
  • N-acetylneuraminic acid is one of the major derivatives of sialic acid. is widely distributed in mammalian cells as the ${\alpha}$2-3- or ${\alpha}$2-6-linked nonreducing terminal residue of oligosaccharide chains of glycoconjugates, and plays important structural and functional roles at the cell membrane surface. The analysis of sialylated glycoproteins is an important part of glycoprotein characterization, especially because sialylation or desialylation in oligosaccharides often causes dramatic changes in the function of glycoproteins. (omitted)

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Glycoproteins isolated from Atractylodes macrocephala Koidz improve protective immune response induction in a mouse model

  • Kim, Kyoung-A.;Son, Young-Ok;Kim, So-Soon;Jang, Yong-Suk;Baek, Young-Hyun;Kim, Chun-Chu;Lee, Jeong-Hoon;Lee, Jeong-Chae
    • Food Science and Biotechnology
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    • v.27 no.6
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    • pp.1823-1831
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    • 2018
  • This study examined the efficacy of Atractylodes macrocephala Koidz (AMK) protein and polysaccharide extracts as adjuvant or adjuvant booster when given together with porcine pleuropneumonia vaccine. Experimental mice (n = 5/group) were subcutaneously immunized with $25{\mu}g$ ApxIIA #3 antigen, a target protein against A. pleuropneumoniae, together with alum and/or various concentrations ($0-500{\mu}g$) of the AMK extracts, while the control group received PBS only. Immunization with ApxIIA #3 antigen increased the antigen-specific IgG titer and this increase was enhanced in the immunization together with AMK protein, but not polysaccharide extract. Supplementation of AMK protein extract exhibited dose-dependent increases in the antigen-induced protective immunity against A. pleuropneumoniae challenge and in the lymphocyte proliferation specific to the antigen. Glycoproteins present in the AMK extract were the active components responsible for immune response induction. Collectively, the present findings suggest that AMK glycoproteins are useful as immune stimulating adjuvant or adjuvant booster.

Efficacy of Brown Seaweed Hot Water Extract Against Hcl-ethanol Induced Gastric Mucosal Injury in Rats

  • Raghavendran Hanumantha Rao Balaji;Sathivel Arumugam;Devaki Thiruvengadam
    • Archives of Pharmacal Research
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    • v.27 no.4
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    • pp.449-453
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    • 2004
  • Effect of pre-treatment with hot water extract of marine brown alga Sargassum polycystum C.Ag. (100 mg/kg body wt, orally for period of 15 days) on HCI-ethanol (150 mM of HCI-etha-not mixture containing 0.15 N HCI in $70\%$ v/v ethanol given orally) induced gastric mucosal injury in rats was examined with respect to lipid peroxides, antioxidant enzyme status, acid/pepsin and glycoproteins in the gastric mucosa. The levels of lipid peroxides of gastric mucosa and volume, acidity of the gastric juice were increased with decreased levels of antioxidant enzymes and glycoproteins were observed in HCI-ethanol induced rats. The rats pre-treated with seaweed extract prior to HCI-ethanol induction reversed the depleted levels of antioxidant enzymes and reduced the elevated levels of lipid peroxides when compared with HCI-ethanol induced rats. The levels of glycoproteins and alterations in the gastric juice were also maintained at near normal levels in rats pre-treated with seaweed extract. The rats given seaweed extract alone did not show any toxicity, which was confirmed by histopathological studies. These results suggest that the seaweed extract contains some anti-ulcer agents, which may maintain the volume/acidity of gastric juice and improve the gastric mucosa antioxidant defense system against HCI-ethanol induced gastric mucosal injury in rats.

Surface glycoproteins determine the feature of the 2009 pandemic H1N1 virus

  • Kim, Jin Il;Lee, Ilseob;Park, Sehee;Park, Man-Seong
    • BMB Reports
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    • v.45 no.11
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    • pp.653-658
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    • 2012
  • After the outbreak of the swine-origin influenza A H1N1 virus in April 2009, World Health Organization declared this novel H1N1 virus as the first pandemic influenza virus (2009 pH1N1) of the $21^{st}$ century. To elucidate the characteristics of 2009 pH1N1, the growth properties of A/Korea/01/09 (K/09) was analyzed in cells. Interestingly, the maximal titer of K/09 was higher than that of a seasonal H1N1 virus isolated in Korea 2008 (S/08) though the RNP complex of K/09 was less competent than that of S/08. In addition, the NS1 protein of K/09 was determined as a weak interferon antagonist as compared to that of S/08. Thus, in order to confine genetic determinants of K/09, activities of two major surface glycoproteins were analyzed. Interestingly, K/09 possesses highly reactive NA proteins and weak HA cell-binding avidity. These findings suggest that the surface glycoproteins might be a key factor in the features of 2009 pH1N1.

Neutral and Amino Sugars Composition of a Lectin from Maackia fauriei (Maackia fauriei 유래 렉틴의 중성당 및 아미노당 조성)

  • Na, Kwang-Heum;Park, Byung-Tae;Park, Jae-Wan;Han, Kyong-Jin;Park, Hyun-Joo;Kim, Ha-Hyung
    • YAKHAK HOEJI
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    • v.53 no.1
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    • pp.34-40
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    • 2009
  • The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Analyzing the composition of monosaccharides, such as that of neutral and amino sugars, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, neutral and amino sugars of lectin obtained from Maackia fauriei were analyzed using an enzyme-linked lectinsorbent assay (ELLA) and high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Peroxidase-labeled lectins such as concanavalin A, Ricinus communis agglutinin, and soybean agglutinin were used for ELLA, since they specifically bind to the monosaccharide residue most frequently encountered in a glycan. The hydrosylate of lectin was prepared by treatment with trifluoroacetic acid, which resulted in the lectin mainly possessing the N-glycan consisting of 98.1 pmol Fuc, 342.1 pmol GlcN, 51.9 pmol Gal, 678.9 pmol Man, and 330.7 pmol Xyl. The present results demonstrate that ELLA and HPAEC-PAD are very effective methods for rapidly estimating the types and relative amounts of monosaccharides in intact glycoproteins.

Detection of Glycoproteins (B and D) and Thymidine Kinase Genes of Herpes simplex virus Type 2 Strain G

  • Kang, Hyun;Park, Jong-Kuk;Uh, Hong-Sun;Kim, Soo-Young;Lee, Hyung-Hoan
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.99-105
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    • 1999
  • BamHI restriction pattern and genomic library of Herpes simplex virus type 2 (HSV-2) strain G were constructed, and locations of the glycoproteins gB and gD, and tk genes on the fragments were detected by Southern blot analysis. HSV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in pHLA2-21 and pHLA2-22 recombinant plasmids, gB gene in pHLA2-24 plasmid, and tk gene in pHLA2-11 clone by Southern blot analysis.

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Oligomerization of the substitution mutants of autographa californica nuclear polyhedrosis Virus (AcNPV) gp64 glycoprotein

  • Kim, Ki-Nam;Poo, Ha-Ryoung;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.72-77
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    • 1997
  • The baculovirus gp64 glycoprotein is a major component of the envelope protein of budded virus (BV). It has been shown that the gp64 glycoprotein plays an essential role in the infection process, especialy fusion between virus envelope and cellular endosomic membrane. Recently we reported optimal conditions required for gp64-mediated membrane fusion in pGP64 DNA transfected Spodoptera frugiperda (Sf9) cells (H. J. Kim and J. M. Yang, Jour, Microbiology, 34.7-14). In order to investigate the role of hydrophobicity within the fusion domain of the gp64 glycoprotein for membrane fusion, 13 mutants which have substitution mutation within hydrophobic region I were constructed by PCR-derived site-derected mutagenesis. Each mutated gp64 glycoproteins was transiently expressed by transfecting plasmid DNA into Spodoptera frugiperda (Sf9) cells. Oligomerization of the transisently expressed gp64 glycoproteins was a nalysed by running them on SDS-polyacrylamide gel electrophoresis under non-reducing condition followed by immunoblotting. All of the mutant gp64 glycoproteins expect cysteine-228 were able to form trimers. These results suggest that hydrophobic region I of the gp64 may not be responsible for the oligomerization of the gp64 glycoprotein.

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