• Korean Journal of Food Science and Technology
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• v.21 no.5
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• pp.606-612
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• 1989

A New lectin from baby clam, Tapes japonica, was isolated and purified through the following procedures; acetone powder, 0.15M NaCl extraction, ammonium sulfate fractionation, N-acetyl-D-galactosamine-agarose affinity column, and ion exchange Mono Q of FPLC. This lectin nonspecifically agglutinated human erythrocytes but didn't agglutinate mouse and rabbit erythrocytes. And the lectin neither stimulated human lymphocytes nor agglutinated Sarcoma 180 cells. On polyacrylamide gel electrophoresis, the lectin migrated as a major single band indicating homogeneous. A molecular weight was estimated to be about 131,000 daltons by Biogel P-300 and 125,000 daltons by SDS-PAGE without ${\beta}-mercaptoethanol$. This lectin is supposed to be a tetramer composed of heterogeneous subunits, about 30,000 and 33,000 daltons. Baby clam lectin was inhibited by EDTA and recovered agglutinating activity by $Ca^{++}\;and\;Mn^{++}$. This lectin is revealed as glycoprotein that contained about 4.2% neutral sugar.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.225-234
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• 2006

In the present study toxicity and immunostimulating activity of the lectin(KML-C), which was extracted from Korean mistletoe(Viscum album coloratum) were investigated in swine. To determine the toxicity, lectin was injected into thigh or cervical muscles of 4-week-old piglets(Landrace) and observed clinically and pathologically. For determination of the immnunostimulating activity, lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine of Aujeszky's disease virus(ADV)(NYJ1-87) which was inactivated by 0.2% formalin was injected into the cervical muscle of antibody-negative piglets in the same age group. Subpopulation of the immune cells and serum neutralizing(SN) antibodies in the piglets were examined after vaccination, and resistance of the piglets against challenge by virulent NYJ1-87 was further examined. The results were also compared with those from piglets injected with aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccine of inactivated NYJ1-87 and NYJ1-87 vaccine without adjuvant, and the results are as follows. By injection of lectin with $30{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets died after signs such as dyspnea, fever, systemic erythema and subcutaneous hemorrhages, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with $7{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets showed signs such as edema and cutaneous hemorrhage in the injected area, lameness and depression, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with 1, 3 and $5{\mu}g/kg$ of body weight to the thigh muscle of each one piglet, signs such as congestion, induration and grayish coloration in the injected area, depression and inappetence were observed in all piglets. Toxic changes were also observed in the liver and kidney of piglets by lectin of 3 and $5{\mu}g$. By injection of lectin with 0.5 and $0.7{\mu}g/kg$ of body weight to the cervical muscle of each 9 piglets, all piglets were clinically normal and there were no significant changes in blood counts and chemistry values. Whereas, epithelial swelling and vacuolation of convoluted tubules were observed from one piglet injected with lectin of $0.7{\mu}g$, and necrosis and fibrosis of muscular fiber were observed in the muscle of one piglet injected with lectin of $0.5{\mu}g$. Only population of sIgM+ B lymphocytes increased among immune cells in all of 15 piglets immunized with lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine, while compared to those in $Al(OH)_3$-adjuvanted vaccine and vaccine without adjuvant. No additional stimulation to the immune cells was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. In piglets immunized with lectin-adjuvanted vaccine, SN titers in reciprocal values for loge were 1.3-4.0 at 1-4 weeks after vaccination, which was similar to those with 1.0-3.3 by vaccine without adjuvant but lower than those with 2.0-5.7 by $Al(OH)_3$-adjuvanted vaccine. Also, no additional increase in the SN titers was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. Piglets immunized with lectin-adjuvanted vaccine were resistant to challenge by the virulent NYJ1-87 at 4 weeks after vaccination, and the SN titers reached to 5.0 one week after challenge, which was higher than those with 4.0 by vaccine without adjuvant but somewhat lower than those with 7.7 by $Al(OH)_3$-adjuvanted vaccine.

• YAKHAK HOEJI
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• v.50 no.3
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• pp.166-171
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• 2006

A calcium-dependent sialic acid-binding lectin has been isolated by thyroglobulin-affinity chromatography from the coastal crab Macrophthalmus Japonicus. This lectin, Macrophthalmus Japonicus lectin (MJL), was eluted with 50mM Tris-HCl, 0.3 M NaCl, 10 mM EDTA, and the recovery yield from the crude protein extract was about 5.6%. The molecular weight of MJL was estimated as 65 kDa in SDS-PAGE both under reducing and non-reducing conditions. MJL induced an agglutination reaction in rabbit, rat, and mouse erythrocytes, but not in human ABO types. This activity was effectively inhibited by sialoglycoproteins such as fetuin, bovine submaxillary mucin, and thyroglobulin.

• Biomolecules & Therapeutics
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• v.18 no.4
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• pp.411-419
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• 2010

Korean mistletoe has a variety of biological effects, such as immunoadjuvant activities. This study investigates the effects of Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) on human T lymphocytes to determine whether VCA acts as an immunomodulator. Purified human T-lymphocytes were cultured with VCA and RNA from each point was analyzed using Affymetrix human genome chips containing 22,500 probe sets which represents more than 18,000 transcripts derived from 14,500 human genes. As a result, there was a striking upregulation of genes coding for chemokines. Seventeen genes out of 50 coding for proteins with chemokine activity were upregulated including CXCL9 and IL-8 which are related to the treatment of cancer. In addition, 28 cytokine genes were upregulated including IL-1, IL-6, IL-8, IFN-$\gamma$, and TNF-$\alpha$. Taken together, the data suggest that Korean mistletoe lectin, in parallel with European mistletoe, has an ability to modulate human T cell function.

• BMB Reports
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• v.36 no.2
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

• Archives of Pharmacal Research
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• v.14 no.3
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• pp.207-216
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• 1991

Pseudixus japonicus agglutinin (PJA) was isolated. And its characteristics were compared with those of concanavalin A (Con A). PJA is a glycopritein composed of 49.3% carbohydrate and 50.7% protein which had relatively high percentages of glutamic acid, aspartic acid and phenylalanine residues. The hemagglutinating activity of PJA was approximately one-eighth of that of Con A when tested with mouse crythrocytes. PJA failed to simulate the proliferation or transformation of human and mouse lymphocytes in contratst to Con A. PJA and Con A showed cytotoxicities against SNU-1 (human stomach cancer cells), SNU-CI (human colon cancer cells) and mouse Sarcoma 180 cells when tested by 3-(4, 5-dimethyl thiazol-2-yl)2. 5-diphenyl tetrazolium bromide (MIT) colorimetric assay. The antitumor activity of the lectin in vivo was also tested in Sarcoma 180 bearing mice. There was no significant difference in prologation of lifc span of the mice after the treatment with PJA and Con A for 10 consecutive days.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.530-530
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• 2000

The host defense system or immune system of all modern animals has their roots in very ancient organisms. After analyzing literature data concerning properties of invertebrates and vertebrates lectins we suggest that mechanism of mannans recognition may exist in marine invertebrates, as a universal mechanism for homeostasis maintenance and host defense, and mannan-binding lectins family of vertebrates has ancient precursor, as was shown for another S-type lectins family. We carried out the screening of mannan-binding type lectin among different species of echinoderms inhabiting in Piter the Grate Bay, the sea of Japan. As a result, the C-type lectins (SJL-32) specific for high mannose glycans was isolated from the coelomic plasma of the sea cucumbers Stichopus japonicus by ion-exchange chromatography on a DEAE-Toyopearl 650M, affinity chromatography on a mannan-Sepharose 6B and gel filtration on a Sephacryl S-200. SJL-32 is homodimer with molecular mass about 32 kDa on SDS-PAGE under non-reducing conditions. Protein part of the lectin has high conteins Asn, Glu, Ser. Hemagglutination of trypsin-treated O blood group human erythrocytes by SJL-32 was competitively inhibited by high-branched -D-mannan composed of -1,2 and -1,6 linked D-mannopyranose residues. In contrast, a variety of mono-, oligo-, and polysaccharides composed of residues of galactose and fucose showed absence or little inhibitory activities. The lectin activity strong depends on Ca2+ concentration, temperature and pH. Monospecific polyclonal antibodies were obtained to the lectin. As was shown by ELISA assay, antibodies to SJL-32 cross-reacted with human serum mannan-binding lectin. This data allows making conclusion about common antigenic determinants and structural homology of both lectins. In our opinion, SJL-32 belongs to evolutionary high conservative mannan-binding lectins (MBLs) family and takes part in the host defense against pathogenic microorganisms.

• 한국당과학회:학술대회논문집
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• 2006.11a
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• pp.63-63
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• 2006

• The Korean Journal of Food And Nutrition
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• v.28 no.6
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• pp.1090-1097
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• 2015

Traditionally, mistletoe is known as an effective anti-cancer medicinal plant, and lectin is recognized as a major component with cytotoxic and immuno-stimulant activity in mistletoe. A Korean mistletoe lectin (KML) has specificity to galactose and galactosamine and is distinguish from European mistletoe lectin (EML). When we examined the concentration of lectin in mistletoe originated from five different types of host trees, the result indicate that the lectin concentration is variable depending on the host tree. Noticeably, mistletoe from chestnut tree contains ten folds higher lectins than that of an oak tree. We also tested the concentration of KML and crude extract (KM-110) of Korean mistletoe that shows 90% cytotoxicity in L5178Y-ML25 lymphoma cell. In addition, the cells show 90% and 70% viability by the treatment of two neutralizing antibodies of KML, 9H7-D10 and 8B11-2C5 neutralization effect with two monoclonal antibodies of KML, 9H7-D10 and 8B11-2C5. Therefore, the result expected that the mistletoe contain some other cytotoxic components except lectin. Finally, the production of $TNF-{\alpha}$ and IL-6 by RAW 264.7 cells stimulated with lectin free-crude extract (LFKM-110) following neutralization by 9H7-D10 monoclonal antibody shows higher than that of lectin containing-crude extract (KM-110). These results suggest that the Korean mistletoe lectin ha a great potential to be developed as therapeutic agent of cancer.

• BMB Reports
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• v.37 no.6
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• pp.715-719
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• 2004

The fate of Telfairia occidentalis seed Agglutinin, TOA, has been monitored during germination. While the level of the Agglutinin in the cotyledons decreased sharply in the first three days to about half of the initial level, it stabilises at this level for the following twelve days. In this interval, Agglutinin activity becomes manifest in the radicle on the fourth day, peaking on the fifth and decreasing rapidly thereafter. In the plumule, the lectin activity becomes manifest on the sixth day, peaks on the seventh and decreases rapidly thereafter. No lectin activity is detectable in any plant tissue including the rump of the cotyledons twenty-seven days after germination. The implications of these observations on the possible role of lectins in plants are discussed.