• Title/Summary/Keyword: Lectins

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The Effects of EGEE on Lectins Binding Pattern of Rat Cerebral Cortex in Developmental Phase (발생기 흰쥐 대뇌 피질의 복합당질에 미치는 ethylene glycol monoethyl ether의 영향)

  • Lee, Eung-Hee;Jo, Un-Bock
    • Journal of Life Science
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    • v.16 no.6
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    • pp.1014-1028
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    • 2006
  • This study attempted to investigate the developmental alterations of the cerebral cortex. The effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by lectin histochemical methods. To investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, Con A and LCA) were detected with by IHC using ABC kit. In the case of injection of EGEE, the reactions to Con A and LCA were shown in binding phase in the cerebral cortex commonly, and the reactions to PNA, RCA-1 and LCA were shown partially, the number of lectins to be shown reaction were decreased, and there were no reactions to DBA, SBA, BSL-1, RCA-1 and UEA-1. The reaction to Con A was similar to control group during developmental phases. The reaction to LCA was increased in the fetal, suckling, and weanning phases compared with control group. But there were no reactions to SBA and sWGA, the reaction to PNA was decreased in the frontal and occipital cortex and no reaction to sWGA in the fetal phase. There were no reactions to sWGA and PNA in the suckling phase and, no reaction to PNA and sWGA. The reaction to Con A was decreased in the frontal, parietal and occipital cortexes and, the reaction to LCA was decreased in the frontal and occipital cortexes in adult phase. As the results, the effects of EGEE, environmental hormone on the each part of cerebral cortex have shown differences. But, It had deep effect on the differentiation and growth in the cerebral cortex pathologically. In particular, the effect was severe in suckling phase. $Galactosyl-({\beta}-1,3)-N-acetyl-D-galactosamine$,${\beta}-N-acetyl-D-galactosamine$ and ${\beta}-N-acetyl-D-glucosamine$ were decreased while ${\alpha}-D-mannose$ and ${\alpha}-D-glucose$ were increased. It affected the sugar metabloism, and it was severe in fetal and suckling phases.

Histological Study of Rat Olfactory Mucosa following Inhalation of Xylene (자일렌이 흡입된 흰쥐 후각점막에 대한 조직학적 연구)

  • Moon, Yong-Suk
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1081-1091
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    • 2018
  • The purpose of this study was to investigate the structure and the glycoconjugate properties of the olfactory mucosa in the rat after inhalation of xylene. Sprague-Dawley male rats were inhalated 300 ppm xylene for 5 times with 5 hours exposure. The olfactory mucosa in the nasal cavity was taken from the animals on 5, 10, 15, 20, and 30 days after inhalation of xylene. The properties of the glycoconjugates in the olfactory mucosa were investigated using 10 biotinylated lectins: (PSA, UEA I, PHA-L, GSL I B4, GSL I, PNA, ECL, SBA, GSL II, and sWGA). In the experimental groups, degenerative changes of the olfactory epithelium were observed until 20 days after inhalation of xylene. In the control group, the olfactory cells in the olfactory epithelium reacted with PSA, UEA I, PNA, SBA, and sWGA, the supporting cells reacted with PSA, PHA-L, GSL I, PNA, ECL, SBA, GSL II, and sWGA, and the Bowman's glands reacted with all 10 lectins. In the experimental groups, the reactivity to PSA, PNA, and SBA in the olfactory cells were decreased, and the reactivity to PSA, PNA, SBA, and GSL II in the supporting cells were decreased. And in the Bowman's gland, the reactivity to PSA, UEA I, GSL I, and sWGA were decreased. Conclusively, the olfactory mucosa was shown a lot of changes in the structure through degenerative process and in the properties of the glycoconjugates after inhalation of xylene. These results suggest that the sugar residues of the glycoconjugates in the olfactory mucosa can be changed by xylene inhalation.

Antifungal Activity and Biochemical Characterization of Lectin Isolated from Locular Fluid of Cherry Tomato Fruit (방울토마토 열매 locular fluid lectin의 항균성과 생화학적 특성)

  • Roh, Kwang-Soo
    • KSBB Journal
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    • v.25 no.3
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    • pp.289-296
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    • 2010
  • Lectins are carbohydrate-binding and a cell-agglutinating proteins, and are concerted with a plants defence mechanism. In particular, chitin-binding lectins in locular fluid of cherry tomato fruit seemed to have a role in defending plants against fungi. The antifungal activity using lectin isolated from locular fluid of cherry tomato fruit was measured in the plant pathogen Cladosporium cucumerinum, Monosporascus cannonballus, Fusarium oxysporum, and Rhizoctonia solani. Amoung the four strains, a potent antifungal activity was detected in Cladosporium cucumerinum and Monosporascus cannonballus, not in Fusarium oxysporum, and Rhizoctonia solani. The molecular weight of this lectin isolated as double protein bands by SDS-PAGE was calculated to be 87 kDa and 47 kDa from the relative mobilities compared with those of reference molecular weight markers. The isolated lectin agglutinated human red blood cells (A, B, AB, O) treated with trypsin, and the most activity was found at B. The optimal temperature of isolated lectin was at $30^{\circ}C$. For the thermal stability, lectin was stable at $20-80^{\circ}C$. The optimal pH of this lectin was at 7.2, and showed complete loss below pH 9.0.

Immune-triggering effect of the foodborne parasite Kudoa septempunctata through the C-type lectin Mincle in HT29 cells

  • Shin, Ji-Hun;Yang, Jung-Pyo;Seo, Seung-Hwan;Kim, Sang-Gyun;Kim, Eun-Min;Ham, Do-Won;Shin, Eun-Hee
    • BMB Reports
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    • v.53 no.9
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    • pp.478-483
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    • 2020
  • Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

Cytokine Expressions with Lectins from Allomyrina dichotoma and Bombyx mori (장수풍뎅이(Allomyrina dichotoma)와 누에(Bombyx mori) 렉틴의 사이토카인 생성 양상 비교)

  • Kim, Se-Jin;Chung, See-Ryun;Jeune, Kyung-Hee
    • Korean Journal of Pharmacognosy
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    • v.36 no.2 s.141
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    • pp.129-135
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    • 2005
  • Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

An EST survey of genes expressed in liver of rock bream(Oplegnathus fasciatus) with particular interests on the stress-responsive and immune-related genes

  • Park, Byul-Nim;Park, Ji-Eun;Kim, Ki-Hong;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.43-43
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    • 2003
  • EST analysis was performed to identify stress-responsive and immune-related genes from rock bream (Oplegnathus fasciatus). cDNA libraries were constructed with liver and randomly chosen 624 clones were subjected to automated sequence analysis. Of 624 clones sequenced in total, approximately 15% of ESTs was novel sequences (no match to GenBank) or sequences with high homology to hypothetical/unknown genes. The bioinforamtic sequence analysis including functional clustering, homology grouping, contig assembly with electronic northern and organism matches were carried out. Several potential stress-responsive biomarker and/or immune-related genes were identified in all the tissues examined. It included lectins, ferritins, CP450, proteinase, proteinase inhibitors, anti-oxidant enzymes, various heat-shock proteins, warm temperature acclimation protein, complements, methyltransferase, zinc finger proteins, lysozymes, macrophage maturation associated protein, and others. This information will offer new possibilities as fundamental baseline data for understanding and addressing their molecular mechanism involved in host defense and immune systems of this species.

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Heterometal-Coordinated Monomeric Concanavalin A at pH 7.5 from Canavalia ensiformis

  • Chung, Nam-Jin;Park, Yeo Reum;Lee, Dong-Heon;Oh, Sun-Young;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2241-2244
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    • 2017
  • The structure of concanavalin A (ConA) has been studied intensively owing to its specific interactions with carbohydrates and its heterometal ($Ca^{2+}$ and $Mn^{2+}$) coordination. Most structures from X-ray crystallography have shown ConA as a dimer or tetramer, because the complex formation requires specific crystallization conditions. Here, we reported the monomeric structure of ConA with a resolution of $1.6{\AA}$, which revealed that metal coordination could trigger sugar-binding ability. The calcium coordination residue, Asn14, changed the orientation of carbohydrate-binding residues and biophysical details, including structural information, providing valuable clues for the development and application of detection kits using ConA.

Lectin Activity and Chemical Characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. from Gastrointestinal Mucosa of Growing Pigs

  • Gao, W.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.863-868
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    • 2004
  • Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.

Roles of Galectin-7 in Cancer

  • Kaur, Manpreet;Kaur, Tarnjeet;Kamboj, Sukhdev Singh;Singh, Jatinder
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.455-461
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    • 2016
  • Galectins are ${\beta}$-galactoside binding lectins that contain one or more carbohydrate recognition domains. As a consequence of sugar-binding properties, galectins exhibit a variety of interactions with glycoproteins, thus playing important roles in various pathological processes. A number of studies have shown roles of galectins in cancer. Galectin-7 is a prototype member of the galectin family implicated in epithelial stratification and cell migration. It can act as a potent dual regulator in different types of cancer. Galectin-7 may contribute either to neoplastic transformation and tumour progression through regulation of cell growth, cell cycle, angiogenesis, apoptosis and cell migration or may have a protective effect in cancer depending on the tissue type. A perusal of the literature indicates particular roles of galectin-7 in carcinomas and melanomas, while contributions await greater exploration in other types of cancers including sarcomas and leukemia. This review collectively summarizes available literature on expression and roles of galectin-7 in different cancers.

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.