• Applied Microscopy
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• v.31 no.1
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• pp.49-57
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• 2001

In this study, the distribution of lectin receptors in culutured fibroblast was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope Labeled sections of the culutured fibroblast revealed gold particles specifically distributed on the cytoplasm and cell surface of the fibroblast. Labeling of 24 hours culutured fibroblast was then quantified and compared to that of 72 hours culutured fibroblast. 24 hours culutured fibroblast sections resulted in specific gold particle distribution on the cytoplasmic vesicle of the culutured fibroblast. These results indicate that lectin WGA receptors are located in the cytoplasmic vesicle and cell surface of the 24 hours culutured fibroblast, and on the cell surface of the 72 hours culutured fibroblast. Therefore, the GlcNAc and NeuNAc regions on the cell surface appear to be functionally associated with cell-recognition and protection from other cell of the tissue, and linked with secretion and exocytosis of the fibroblast cytoplasm.

• Journal of Life Science
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• v.23 no.3
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• pp.347-354
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• 2013

Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.

• Applied Microscopy
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• v.30 no.1
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• pp.101-111
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• 2000

In this study, the distribution of lectin receptors in Paragonimus westermani tissue was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope. Labeled sections of the metacercaria revealed gold particles specifically distributed on the tegumental syncytium and lamella of the excretory canal. Labeling of young adult tissue was then quantified and compared to that of adult worm tissue. Adult worm tissue sections resulted in specific gold particle distribution on the lamella of caecal epithelium and excretory canal. These results indicate that lectin WGA receptors are located in the tegumental syncytium and lamella of the excretory canal of the metacercariae, and in the lamella of the caecum and excretory canal of the young adult and adult. Therefore, the GlcNAc and NeuNAc regions in the tegumental syncytium appear to be functionally associated with cell-recognition and protection from the immune system of the host, and linked with membrane transport and absorption of nutrients in the lamella of the excretaory canal and caecal epithelia.

• Journal of Life Science
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• v.33 no.8
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• pp.632-638
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• 2023

A soybean cultivar with a green seed coat and cotyledon contains high levels of lutein, which is beneficial for eye health. Plus, antinutritional components such as lipoxygenase, Kunitz trypsin inhibitor (KTI), lectin and stachyose exist in the mature seed. The genetic elimination of these antinutritional factors is a necessary step in green soybean breeding. This research was conducted to improve a new green soybean line with the green cotyledon and tetra null genotype (lox1lox2lox3tilers2) in terms of lipoxygenase, KTI, lectin and stachyose. We used five germplasms to develop a breeding population. A total of 69 F₂ seeds were obtained from the cross of parent 1 and parent 2, and from those, 21 F₂ seeds were selected that had the green seed coat color, and which were free of lectin protein. Next, four F₂ plants with the green seed coat and tetra null genotype were selected from the breeding population derived from four genotypes. The absence of lipoxygenase, KTI and lectin proteins was confirmed in the F₅ strain. The breeding line has a green seed coat, green cotyledon and white hilum color. The 100-seed weight and stachyose content for the breeding line were 30.7 g and 2.40 g/kg, respectively. The line selected in this study could be used as a cultivar or parent to improve colored soybean cultivars through the removal of antinutritional components such as lip- oxygenase, KTI, lectin and stachyose.

• 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.

• Journal of fish pathology
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• v.25 no.1
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• pp.11-20
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• 2012

C-type lectins are crucial for pathogen recognition, innate immunity, and cell-cell interactions. In this study, a C-type lectin gene was cloned from the rock bream. The full-length RbCTL cDNA was 729 bp with a 429 bp ORF encoding a 164-residue protein. The deduced amino acid sequence of RbCTL had all of the conserved features crucial for its fundamental structure, including the four cysteine residues involved in sulfide bridge formation and potential $Ca^2+$/carbohydrate-binding sites. RbCTL contains a signal peptide one single carbohydrate recognition domain. It showed 29.4% similarity to the C-type lectin of rainbow trout. RbCTL mRNA was predominately expressed in gill and head-kidney tissue and expressed less in peripheral blood leukocytes, trunk-kidney, spleen, liver, intestine and muscle. Expression of RbCTL was differentially upregulated in rock bream stimulated with LPS, Con A/PMA and poly I:C.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.531-539
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• 1996

The lectin-binding patterns in the testis of the sexually matured Jindo dog were investigated to study the distribution of glycoconjugates in the seminiferous tubule under light and transmission electron microscopy. Positive reactions to Wheat germ agglutinin(WGA) and Dolichos biflorus agglutinin (DBA) were observed in the Sertoli cell and in the residual body of spermatid with a stronger reaction in the Sertoli cell to the lectins than in the residual body. Strong reactions to Soybean agglutinin(SBA) and Peanut agglutinin(PNA) were observed in the acrosome vesicles of the Golgi- and cap-phase spermatid, while a moderate reaction was observed in the acrosome-phase, maturation-phase spermatid and the residual body. The acrosome area of the spermatid reacted intensively to Griffonia simplicifolia agglutinin( GS-I) when the cell was in the acrosome-phase and maturation-phase, and the same reaction to the GS-I was observed in the residual body. However, the seminiferous tubule did not react to Ulex europeus agglutinin I(UEA-I). The gold-labelling of the Sertoli cells with DBA resulted in positive reactions of the Sertoli cell column and processes when observed under the electron microscopy, while the Golgi-, cap- and acrosome-phase spermatids reacted positively to SBA in the peripheral low-dense area of the acrosome vesicle of spermatid. Based on these results, we concluded that differences in the lectin-binding pattern of the seminiferous tubules were recognized in the Jindo dog compared to other animals.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.547-554
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• 1997

Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.381-381
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• 2000

Lectin binding assay was conducted on 3 A. tamarense isolates (AT-A, AT-2 and AT-6). Fatty acid composition of all 3 isolates was analyzed, and total carotenoid content and $\beta$-carotene were also determined. AT-A and AT-2 treated with different lectins in this study showed the positive response, whereas potentially toxic AT-6 did not bind DBA lectin, regardless of different growth phase, but conjugated ConA, PNA, RCA, SBA, UEA and WGA. It is possible that DBA is a desirable method for rapid and easy discrimination of highly toxic A. tamarense. AT-A, AT-2 and AT-6 comprised saturated fatty acids (49.0-61.9%), monounsaturated fatty acids (8.0-20.5%) and polyunsaturated fatty acids (23.2-30.5%). In particular, 22:6 (n-3) polyunsaturated fatty acid in AT-6 had a high abundance, compared with AT-A and AT-2. However, carotenoid content and $\beta$-carotene were not contributed to discriminate each isolate. Due to variability in biochemical composition at different isolates, possibly DBA and 22:6 (n-3) polyunsaturate fatty acid provide a good information for discrimination of AT-6.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.43-47
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• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.