• Title/Summary/Keyword: Lectins

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Molecular cloning and expression analysis of a C-type lectin in the rock bream, Oplegnathus fasciatus

  • Kwon, Mun-Gyeong;Kim, Ju-Won;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Park, Chan-Il
    • 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.

Purification and Characterization of a New Galactoside Specific Lectin from Trichosanthes kirilowii Root

  • Yun, Doo-Hee;Park, Eun-Ju;Park, Jong-Ok;Lee, Young-Han;Seo, Jeong-Kon;Ryu, Sung-Ho;Suh, Pann-Ghill;Kim, Hee-Sook
    • BMB Reports
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    • v.28 no.1
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    • pp.6-11
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    • 1995
  • A new lectin, named TRA, was purified from Trichosanthes kirilowii root by acid-treated Sepharose 6B, Mono-Q, and TSK-gel 3000SW column sequential chromatography. The lectin appeared homogeneous by native gel electrophoresis at pH 4.3 and gave two protein bands of Mr=31 and 28 kDa by SDS-PAGE. The N-terminal amino acid sequences of the polypeptides of TRA have not been reported in amino acid sequences of the lectins. TRA lectin formed a precipitate with asialofetuin, neuraminidase-treated fetuin. A sugar inhibition assay indicated that N-acetyl-D-galactosamine, among the monosaccharides tested, was the most potent inhibitor of TRA-induced hemagglutination. Asialofetuin showed a 260-times stronger inhibitory activity than N-acetyl-D-galactosamine. TRA lectin also showed agglutination with normal leukocytes and lymphoma cells, but not with premature hemopoietic cells. These results suggest that TRA is a novel plant lectin.

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Loss of Potential Biomarker Proteins Associated with Abundant Proteins during Abundant Protein Removal in Sample Pretreatment

  • Shin, Jihoon;Lee, Jinwook;Cho, Wonryeon
    • Mass Spectrometry Letters
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    • v.9 no.2
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    • pp.51-55
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    • 2018
  • Capture of non-glycoproteins during lectin affinity chromatography is frequently observed, although it would seem to be anomalous. In actuality, lectin affinity chromatography works at post-translational modification (PTM) sites on a glycoprotein which is not involved in protein-protein interactions (PPIs). In this study, serial affinity column set (SACS) using lectins followed by proteomics methods was used to identify PPI mechanisms of captured proteins in human plasma. MetaCore, STRING, Ingenuity Pathway Analysis (IPA), and IntAct were individually used to elucidate the interactions of the identified abundant proteins and to obtain the corresponding interaction maps. The abundant non-glycoproteins were captured with the binding to the selected glycoproteins. Therefore, depletion process in sample pretreatment for abundant protein removal should be considered with more caution because it may lose precious disease-related low abundant proteins through PPIs of the removed abundant proteins in human plasma during the depletion process in biomarker discovery. Glycoproteins bearing specific glycans are frequently associated with cancer and can be specifically isolated by lectin affinity chromatography. Therefore, SACS using Lycopersicon esculentum lectin (LEL) can also be used to study disease interactomes.

Lectine-binding patterns of spermatogenic cells in the Jindo dog (진도견 정자형성계 세포들의 Lectin-binding patterns)

  • Park, Young-seok;Lee, Seong-ho
    • 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.

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Discrimination of three highly toxic Alexandrium tamarense(Dinophyceae) isolates using FITC-conjugated lectin probesLectin probe

  • Jo Eun Seop;Jo Yong Cheol;Kim Tae Jin;Kim Hak Gyun
    • 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.

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Isolation and Characterization of a Trypsin Inhibitor and a Lectin from Glycine max cv. Large Black Soybean

  • Ye, Xiu Juan;Ng, Tzi Bun
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1173-1179
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    • 2009
  • Trypsin inhibitors and lectins are defense proteins produced by many organisms. From Chinese 'Large Black Soybeans', a 60 kDa lectin and a 20 Da trypsin inhibitor (TI) were isolated using chromatography on Q-Sepharose, Mono Q, and Superdex 75. The TI inhibited trypsin and chymotrypsin with an $IC_{50}$ of 5.7 and $5{\mu}M$, respectively. Trypsin inhibitory activity of the TI was stable from pH 3 to 13 and from 0 to $65^{\circ}C$. Hemagglutinating activity of the lectin was stable from pH 2 to 13 and from 0 to $65^{\circ}C$. The TI was inhibited by dithiothreitol, signifying the importance of disulfide bond. The TI and the lectin inhibited HIV-1 reverse transcriptase ($IC_{50}$=44 and $26{\mu}M$), and proliferation of breast cancer cells ($IC_{50}$=42 and $13.5{\mu}M$) and hepatoma cells ($IC_{50}$=96 and $175{\mu}M$). The hemagglutinating activity of the lectin was inhibited most potently by L-arabinose. Neither the lectin nor the TI displayed antifungal activity.

Korean Mistletoe Lectin-induced Apoptosis in Hepatocarcinoma Cells is Associated with Inhibition of Telomerase via Mitochondrial Controlled Pathway Independent of p53

  • Park, Won-Bong;Lyu, Su-Yun;Choi, Sang-Ho
    • Archives of Pharmacal Research
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    • v.25 no.1
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    • pp.93-101
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    • 2002
  • The extract of European mistletoe ( Viscum album, L) has been used in adjuvant chemotherapy of cancer and mistletoe lectins are considered to be major active components. The present work was performed to investigate the effects of Korean mistletoe lectin (Viscum album L. coleratum agglutinin, VCA) on proliferation and apoptosis of human hepatoma cells as well as the underlying mechamisns for these effects. We showed that VCA induced atoptosis in both SK-Hep-1 and Hep 3B (p53-negative) cells through p53- and p21 -independent pathways. VCA induced apoptosis by down-regulation of Bcl-2 and by up-regulation of Bax functioning upstream of caspase-3 in both cell lines. In addition, we observed down-regulation of telomerase activity in both VCA-treated cells. Our results provide direct evidence of the anti-tumor potential of this biological response which comes from inhibition of telomerase and consequent inducing apoptosis. VCA-induced apoptosis is regulated by mitochondria controlled pathway independently of p53. These findings are important for the therapy with preparation of mistletoe because they show that telomerase-dependent mechanism can be targeted by VCA in human hepatocarcinoma. Taken together, our results suggest that the VCA, considered as a telomerase-inhibitor, can be envisaged as a candidate for enhancing sensitivity of conventional anticancer drugs.

Analysis and Characterization of the Taro (colocasia antiquorum) lsolectin (토란 Isolectin의 분석 및 특성)

  • 서영주;삼호정만
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.308-314
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    • 1994
  • Four Taro isolectins (I, II, III, IV) were purified by ammonium sulfate, chromatography on CM-celluose and isoelectric focusing. I and IV lectins proved homogeneous by disk polyacrylamid gel electrophoresis and densitometric patterns. But in the presence of urea, IV lectin further dissociated into two different subunits. These lectinis had different hemagglutinating activities and inhibition in their activities after mixed with pepsin particuclary, but not with carbohydrates, heating pH, urea, guanidine, trypsin, pronase and $Ca^{2+}$.

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LECTIN BINDING PATTERNS IN LARYNGEAL KERATOSIS WITH ATYPIA (후두각화증에서 이형성 유무에 따른 렉틴 반응)

  • 김광문;김기령;윤주헌;장미숙;조정일
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1991.06a
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    • pp.16-16
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    • 1991
  • 렉틴(Lectins)은 세포막의 당단백질의 과당류 말단기에 특이적으로 결합하는 비면역성의 당단백질 혹은 단백질을 말하는데 정상세포와 변형된 세포에서 렉틴반응의 차이가 남으로써 요즘에는 렉틴을 병리학적 진단에 이용 가능하게 되었다. “Loss of contact inhibition”은 세포 악성화의 중요한 특징으로 세포의 악성화는 세포막의 구조의 변화와 관계가 있을 것으로 알려져 왔으며 1989년 후두암에서 PNA의 반응이 양성 반응을 나타낸다고 보고되어 저자들은 7가지의 렉틴을 이용하여 후두암전구증의 하나인 후두각화증에서 이형성(Atypia)의 존재유무에 따른 렉틴 반응의 변화를 알아보고자 본 연구를 시행하여 다음과 같은 결과를 얻었다. 1. 이형성이 없는 단순 후두각화증에서는 Con A와 WGA만이 기저세포에 반응이 있었으며, 후두각화증의 가장 중요한 부위인 극세포층에서는 WGA, RCA-I, PNA, SBA, UEA-I, DBA가 세포질에는 반응이 없이 세포간교에만 염색이 되는것을 관찰할 수 있었고 Con A는 반대로 극세포층의 세포질에 반응을 하였으나 세포간교에는 반응이 없었다. 2. 이형성을 동반한 후두각화증에서, 기저세포에서는 반응의 차이가 없었고 극세포층에서는 UEA-I, RCA-I. WGA, PNA, SBA는 세포간교에서 반응이 나타나지 않았고 세포질에서 양성 반응을 관찰 할 수 있었으며 DBA, Con-A에서는 반응의 차이를 관찰 할 수 없었다. 따라서 후두각화증의 극세포층 세포막에서 UEA-I, RCA-I, WGA, PNA, SBA 반응의 소실은 후두암으로의 진행과 밀접한 관계가 있는 이형성의 존재를 암시한다하겠다.

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Isolation and Characterization of $\beta$-Galactoside Specific Lectin from Korean Mistletoe (Viscum album var. coloratum with Lactose-BSA-Sepharose 4B and Changes of Lectin Conformation

  • Park, Won-Bong;Ju, Yeun-Jin;Han, Seon-Kyu
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.429-435
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    • 1998
  • Lectins and its A- and B-chains from Korean mistletoe (Viscum album var. coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to .epsilon.-amino groups of lysine residues of spacer (BSA) after reduction by $NaCNBH_3$. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A-chain and a 35kD of B-chain. Amino acid analysis and N-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturaiional red shift which also correspond to the conformations and activity of lectin.

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