• YAKHAK HOEJI
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• v.45 no.2
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• pp.180-189
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• 2001

We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

• Journal of the Korean Wood Science and Technology
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• v.29 no.4
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• pp.79-88
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• 2001

Lectin was isolated from shiitake mushroom (Lentinula edodes) with 0.15 M NaCl solution, and purified by the following procedures : precipitation by ammonium sulfate, anion exchange column chromatography on DEAE Sephadex A-50 and hydroxyapatite column chromatography. The fresh pileus part of the mushroom contained more than two times of lectin compared to the stipe part, and lectins and its activity were reduced by heating. The extraction yield of crude lectin was 46.03%, 28% yield after purification on on DEAE Sephadex A-50 column chromatography. Some amino acids, aspartic acid, serine, alanine and histidine, were increased by purification process. Relatively low molecular weight parts of lectin had the agglutinating activity for rabbit blood, and its molecular weight was about 23 kDa The molecular weights of purified lectins, LA-a and LB-b, by the hydroxyapatite column chromatography were 24 kDa and 23 kDa, respectively.

• KSBB Journal
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• v.24 no.2
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• pp.170-176
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• 2009

Lectin was finally isolated on Sephadex G-100 from Korean soybean cultivars developed for soy source and investigated its some biochemical properties. Native PAGE pattern of this lectin revealed a molecular weight of 108 kDa as tetramer. The molecular weight of this lectin isolated as double protein band by SDS-PAGE was calculated to be 32 and 22 kDa from the relative mobilities compared with those of the standard proteins. Among the tested red blood cell, the isolated lectin agglutinated rabbit red blood cell treated with trypsin, but did not agglutinated human red blood cells (A, B, AB, O), rat, and untreated rabbit red blood cell. The optimal temperature and thermal stability of isolated lectin was at 20-$50^{\circ}C$ and 10-$60^{\circ}C$, respectively. This lectin was stable at 7.2, and showed complete loss in its activity below pH 6.2 and above pH 8.0.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.421-432
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• 1997

A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.

• 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.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.248-261
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• 1994

Background: Lectins are proteins or glycoproteins of non-immune origin that recognize a specific sequence of sugar residues. The availability of a large number of lectins has provided the capacity to identify selectively glycoconjugates possessing distinctive chemical structure in diverse sites of highly specialized biological activity. The purpose of the present study was to investigate the lectin binding patterns of various components in human pulmonary tubercles. Method: Biopsy specimens of tuberculous lung were obtained from male adult patients who underwent a surgical resection for severe pulmonary tuberculosis. The specimens were processed and stained with 13 kinds of biotinylated lectins according to some modification of Hsu and Raine's methods. Results: 1) In the caseous necrotic lesions, BS $I-B_4$ showed negative reaction and BS I were also negative except some irregularly-shaped cells located in the marginal zone. All other lectins, however, showed a positive reaction with various binding patterns. 2) The epithelioid cells were broadly divided into three groups according to the reaction patterns in the cytoplasms and cell membranes. 3) WGA, ECL, PHA-L, PHA-E and LCA showed strong staining in the lymphocytes. 4) SBA showed a different binding patterns between the endothelial layers located in the region beyond the fibrous layers and those located within the fibrous layers. 5) PNA showed a positive reaction in the outer 1/3 to 1/2 of the fibrous layer, but showed no staining in the inner 1/2 to 2/3 of the fibrous layers. Conclusion: The present lectin histochemical study provided a useful information to assess the characterization and distribution of various glycoconjugates in each constituent of human pulmonary tubercles. The results demonstrate structural differences in the glycoconjugate composition of various components of the tubercles and reveal changes in glycosylation in the components during soft tubercle formation. This study provides a new data useful for the studies on the pathogenesis and pathology of human pulmonary tubercles.

• Applied Microscopy
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• v.39 no.4
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• pp.291-299
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• 2009

We report on a quantitative analysis of cone and rod photoreceptors in hamster retina. Cone and rod photoreceptors were counted in retinal whole mounts using differential interference contrast (DIC) optics microscopy after staining of cone photoreceptors were stained with peroxidase-labeled peanut lectin. Middle-to-long-wave-sensitive-(M/L-), and shortwave-sensitive-(S-) cone opsins were visualized by observed using confocal microscope after immunocytochemical procedure. The average cone density was 9,307 $cells/mm^2$, giving a total of cones of 293,060 cone cells per retina. The peak density of cone cells (12,857 $cells/mm^2$) was found 0.3 mm from the optic disk (OD) of the nasal retina. The average rod density was 300,082 $cells/mm^2$, giving a total number of rods of 9,448,150 cells. The peak density of rod cells was found 0.3 mm from the OD of the dorsal retina. Of all photoreceptors studied, the total percentage of rods and cones were 96.99% and cones 3.01%, respectively. The mean ratio of rod and cone was 32.24 : 1. The cone photoreceptors of hamster contained both M/L- and S-cone opsins. The present results suggest that the hamster retina is strongly rod-dominated with some photopic property of vision.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.1-6
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• 2002

Synthesis of new active protein was investigated by heat-treatment and fermentation of kidney beans with Bacillus subtilis ATCC 51189. The amount of water-soluble protein in raw kidney bean (raw protein, RP) was greatly reduced by heating (heated protein, HP) and several new amino acids were synthesized by fermentation. The molecular weights of proteins determined by SDS-PAGE were 118 kDa for RP and a new band of 18.0 kDa For protein (fermented protein, FP) in kidney beans heated and fermented with B. subtilis ATCC 51189. Hemagglutinating activities of RP, HP and FP were 128 HU, 4 HU and 32 HU respectively. Both of RP and FP showed anticancer activity against stomach cancer cell line (SNU-1) at 50 $\mu\textrm{g}$g/mL and lymphocyte stimulating activity at 1 $\mu\textrm{g}$/mL, and stimulated PBMC to secrete IFN-${\gamma}$ and IL-12. However, HP did not show any kinds of activities. Taken together, these results suggested that lectin in kidney beans was destroyed by heating, hut new active lectin-like Protein was derived by fermentation with B. subtilis ATCC 51189.

• Journal of Life Science
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• v.27 no.6
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• pp.632-639
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• 2017

Mature haptoglobin (Hp) is a plasma glycoprotein and acts as an antioxidant by scavenging cell-free hemoglobin (Hb). Prohaptoglobin (proHp) is an unprocessed Hp precursor which is present a little in circulation. However, the biological function of proHp remains unknown. To investigate the structural and functional differences between proHp and Hp, we prepared recombinant proHp isoforms and compared their sialic acid content and Hb-binding capacity with those of mature isoforms. When proHp samples were analyzed by Western blot under non-reducing conditions, proHp1 was detected as one band of approximately 130 kDa and proHp2 as multiple bands >200 kDa, in the manner of mature Hp1-1 and Hp2-2, respectively. On the native polyacrylamide gel under non-reducing and non-denaturing conditions, both proHp isoforms migrated more slowly than their mature Hp counterparts. In addition, the lectin-based ELISA assay demonstrated that the content of sialic acid in proHp1 and proHp2 was much less than in Hp1-1 and Hp2-2. The Hb-binding capacity of proHp was also lower than those of mature Hp. These findings indicate that proHp and Hp are similar in the size and polymerization pattern, but different in sialic acid content and Hb-binding activity. It suggests precursor proHp may exert different functions in circulation than does mature Hp.

• Journal of Life Science
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• v.18 no.11
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• pp.1543-1550
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• 2008

We studied the effects of smoking, which is one of indoor-environmental pollutants and related to various cancers, on glycoconjugates of rat sebaceous glands with the lectin histochemistry. To investigate the effects of smoking on glycoconjugates, male Sprague-Dawley rats were exposed to tabacco smoke for 10 minutes per day in an inhalation chamber for 1, 2, 3, and 5 days with active and passive exposure. For the structure of sebaceous glands we used PAS reaction, and for the glycoconjugates binding pattern 9 biotinylated lectins (DBA, SBA, PNA, BSL-1, WGA, RCA-1, UEA-1, Con A, and LCA) were used. Some remarkable changes, such as the decrease in the size of sebaceous glandular acini, the destruction of upper portion of sebaceous glands, vacuolation of central portion of sebocytes, and the immature sebaceous glandular acini were seen in the smoke-exposed rats. In the control rats, basal cells were stained with BSL-1, PNA and WGA, but the stronger reaction was founded in BSL-1 binding. Also, sebocytes were stained with PNA, WGA, Con A, BSL-1 and SBA, but stronger reactions were founded in PNA and Con A stainings. Specific changes in the lectin binding patterns were also observed in the smoke-exposed rats. In the basal cells of exposed rats, PNA binding increased, BSL-1 decreased but returned to control level, and WGA disappeared. Plus, immature glandular acini, which were not found in the control rats, were stained PNA, Con A and BSL-1, but the stronger reaction were founded in PNA and Con A binding. In conclusion, it was assumed that the tabacco smoke seriously effected on the structure and glycoconjugates metabolism of sebaceous glands.