• 제목/요약/키워드: Glycoproteins

검색결과 209건 처리시간 0.034초

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
    • /
    • 제5권4호
    • /
    • pp.227-233
    • /
    • 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.

  • PDF

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

  • Shin, Jihoon;Cho, Wonryeon
    • Mass Spectrometry Letters
    • /
    • 제7권3호
    • /
    • pp.74-78
    • /
    • 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.

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

  • Kang, Man-Sik;Song, Woo-Keun;Ho. Woo Nam;Chung, Chang-Yong
    • 한국동물학회지
    • /
    • 제28권3호
    • /
    • pp.125-136
    • /
    • 1985
  • 본 연구는 계배근세포가 분화하는 과정에서 조절을 받는 단백질이 있는 지의 여부를 가려내기 위해서 근세포 및 그 원형질막의 당단백질의 변화양상을 표지된 Con A 염색법을 써서 검토한 것이다. 당단백질 중에는 세포에서만 발견되는 것이 8종, 원형질막에서만 발견되는 것이 4종, 그리고 공통적으로 발견되는 것이 9종이 있었다. 그중에서 분화하는 동안에 변하지 않는 것, 증가하는 것, 감소하는 것, 증가하다 감소하는 것, 그리고 감소하다 증가하는 것 등 다섯가지 종류가 있었다. 본 연구에서 fibronectin이 근관형성후에는 감소되는 사실이 판명되었는데, 이 결과는 지금까지 논란의 대상이 되어오던 상반된 결과들이 분화과정 중에서 택한 시기의 차이에 따라 나타난 결과들로 볼 수 있음을 보여주었다. 일반적으로 근세포의 융합이 진행될수록 고분자량의 당단백질은 감소하고, 대신 저분자량의 당단백질은 증가하는 경향성을 보였고, 이와같은 결과는 근원세포가 융합을 함에 따라서 당단백질의 구조적인 재편성이 일어남을 암시하는 것으로 받아들여졌다.

  • PDF

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

  • Cho, Due-Hyeon;Ryu, Chang-Soo;Park, Jee-Hun;Kim, Ha-Hyung
    • 대한약학회:학술대회논문집
    • /
    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
    • /
    • pp.316.1-316.1
    • /
    • 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)

  • PDF

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
    • /
    • 제27권6호
    • /
    • pp.1823-1831
    • /
    • 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
    • /
    • 제27권4호
    • /
    • pp.449-453
    • /
    • 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
    • /
    • 제45권11호
    • /
    • pp.653-658
    • /
    • 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.

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

  • 나광흠;박병태;박재완;한경진;박현주;김하형
    • 약학회지
    • /
    • 제53권1호
    • /
    • pp.34-40
    • /
    • 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
    • 대한바이러스학회지
    • /
    • 제29권2호
    • /
    • pp.99-105
    • /
    • 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.

  • PDF

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
    • /
    • 제35권1호
    • /
    • pp.72-77
    • /
    • 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.

  • PDF