• 제목/요약/키워드: N-glycoprotein

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Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines

  • Talabnin, Krajang;Talabnin, Chutima;Ishihara, Mayumi;Azadi, Parastoo;Wongkham, Sopit;Sripa, Banchob
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권2호
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    • pp.691-695
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    • 2016
  • Protein glycosylation is the most common posttranslational modification in mammalian cells. Aberrant protein glycosylation has been reported in various diseases, including cancer. We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-$MS^n$). Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by $NeuA_{c2}Gal_1GalNA_{c1}$. These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA.

해산 극피동물 중의 당단백질의 특성과 이용 I. 해삼 당단백질 및 황산콘드로이친의 화학조성과 특성 (Chemical Compositions of Glycoprotein and Chondroitin Sulfates from Sea Cucumber(Stichopus japonicus))

  • 류홍수;문정혜;서재수
    • 한국식품영양과학회지
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    • 제26권1호
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    • pp.72-80
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    • 1997
  • 식용 및 약용으로 이용한 역사가 오래인데도 불구하고 구성성분의 조성에 대하여 충분한 연구가 되어 있지 않은 해삼에서 당단백질과 황산콘드로이친을 추출정량하여 이들의 화학적 조성과 구조적 특성을 GC. MS및 IR로 검토하였다. 동결건조 해삼분말의 화학적인 조성은 sulfate esters함량이 0.90~l.21%, 다당류 함량이 23.08~26.97%였고 이를 구성하고 있는 구성단당류는 fucose의 함량이 30%이상을 차지하는 것을 비롯하여 glucose, N-acetylglucosamine, mannuronic acid, ribose의 5종이 거의 88%이상을 차지하고 있었고 그외 mannose, galactose, N-acetylgalactosaminne, D-glucuronic acid도 소량 검출되었다. 추출당단백질의 sulfate esters의 함량은 0.96~l.13%, 다당류의 함량은 58.7%~62.7%로 나타나고 주요 구성당은 fucose가 30%내외로 가장 많았으며 mannose, glucose와 ribose가 86%이상을 차지하며 galactose, N-acetylgalactosamine, D-glucuronic acid, N-acetylglucosamine도 소량 함유하고 있었다. 동결건조 해삼분말에서 추출한 황산콘드이친의 황산기의 함량은 3.52~3.72%이었다. 이들의 구성성분은 fucose, N-acetylgalactosamine 및 D-glucuronic acid 이며 IR spectrum에서 1240$cm^{-1}$ /의 S=0신축진동과 850$cm^{-1}$ /나 820$cm^{-1}$ /의 C-O-S 신축진동이 나타나 fucose의 $C_4$의 결합위치를 나타내고 있었다.

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Sialylated oligosaccharide analyses using high-performance liquid chromatography with a fluorescence detector

  • Cho, Due-Hyeon;Ryu, Chang-Soo;Park, Jee-Hun;Kim, Ha-Hyung
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.316.1-316.1
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    • 2003
  • N-acetylneuraminic acid is one of the major derivatives of sialic acid. is widely distributed in mammalian cells as the ${\alpha}$2-3- or ${\alpha}$2-6-linked nonreducing terminal residue of oligosaccharide chains of glycoconjugates, and plays important structural and functional roles at the cell membrane surface. The analysis of sialylated glycoproteins is an important part of glycoprotein characterization, especially because sialylation or desialylation in oligosaccharides often causes dramatic changes in the function of glycoproteins. (omitted)

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hEPO 유전자의 발현이 조절되는 형질전환 닭의 생산 (Generation of Transgenic Chickens Regulating hEPO Gene Expression)

  • 구본철;권모선;김태완
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.193-199
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    • 2010
  • We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

락토페린의 최근 연구 개발 동향 (Current Trends in Lactoferrin Research and Development)

  • 류연경;김완섭
    • Journal of Dairy Science and Biotechnology
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    • 제27권1호
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    • pp.19-28
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    • 2009
  • Lactoferrin was first identified 60 years ago as a "red protein" in bovine milk. Lactoferrin, one of the transferrin family proteins, is an iron-binding glycoprotein found in milk and various mucosal secretions; it is also released from activated neutrophils. Human lactoferrin has a molecular weight of 82.4 kDa and is composed of 702 or 692 amino acid residues. Bovine lactoferrin has a molecular weight of 83.1 kDa and is composed of 689 amino acid residues. Both lactoferrin and transferrin have the ability to bind two $Fe^{3+}$ ions, together with two ${CO_3}^{2-}$ ions with extremely high affinity; these proteins also have the ability to release this iron at low pH levels. The polypeptide chain in lactoferrin is folded into two globular lobes, representing the N-terminal and C-terminal halves. Both lobes have similar folding and 40% sequence identity. This protein is capable of multiple functions as described in various review papers, including antimicrobial, antiviral, antiinflammatory, anticancer, antioxidant, and cell growth-promoting activities. Lactoferrin also exhibits immunomodulating effects and plays an active role in the regulation of myelopoiesis and the inhibition of bacterial translocation.

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Characteristics of a Low Molecular Weight Minor Anionic Isoperoxidase $A_{3n}$ from Radish

  • Lee, Mi-Young;Kim, Soung-Soo
    • BMB Reports
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    • 제31권6호
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    • pp.548-553
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    • 1998
  • A minor anionic isoperoxidase named $A_{3n}$, was isolated from Korean radish (Raphanus sativus L.) root. Purification of the enzyme was accomplished by CMcellulose chromatography, DEAE-Sephacel chromatography, and Sephadex G-75 gel filtration. The enzyme was a glycoprotein with molecular weight of approximately 31,000 as determined by SDS-PAGE and 33,000 by Sepadex G-150 gel filtration, which is by far the smallest among the reported isoperoxidases. The pI value was 3.5. The optimum pH of the enzyme was 6.5 for guaiacol and $H_2O_2$, and the $K_m$ values for guaiacol and $H_2O_2$ were 13.3 mM and 1.5 mM, respectively. Kinetic studies with various substrates revealed that only A3n, unlike other isoperoxidases from radish, did not use scopoletin as a substrate and had very low $K_m$ value of 0.25 mM for ferolic acid among naturally occurring phenolic substrates.

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식물의 자가불화합성, 최근의 진보 (Recent Advances in the Studies of Self-Incompatibility of plants)

  • 한창열;한지학
    • 식물조직배양학회지
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    • 제21권5호
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    • pp.253-275
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    • 1994
  • Many flowering plants possess genetically controlled self -incompatibility (SI) system that prevents inbreeding and promotes outcrosses. SI is usually controlled by a single, multiallelic S-locus. In gametophytically controlled system, SI results when the S-allele of the pollen is matched by one of the two S-alleles in the style, while in the sporophytic system self-incompatible reaction occurs by the interaction between the pistil genotype and genotype of, not the pollen, but the pollen parent In the former system the self-incompatible phenotype of pollen is determined by the haploid genome of the pollen itself but in the latter the pollen phenotype is governed by the genotype of the pollen parent along with the occurrence of either to-dominant or dominant/recessive allelic interactions. In the sporophytic type the inhibition reaction occurs within minutes following pollen-stigma contact, the incompatible pollen grains usually failing to germinate, whereas in gametophytic system pollen tube inhibition takes place during growth in the transmitting tissue of the style. Recognition and rejection of self pollen are the result of interaction between the S-locus protein in the pistil and the pollen protein. In the gametophytic SI the S-associated glycoprotein which is similar to the fungal ribonuclease in structure and function are localized at the intercellular matrix in the transmitting tissue of the style, with the highest concentration in the collar of the stigma, while in the sporophytic SI deposit of abundant S-locus specific glycoprotein (SLSG).is detected in the cell wall of stigmatic papillae of the open flowers. In the gametophytic system S-gene is expressed mostly at the stigmatic collar the upper third of the style length and in the pollen after meiosis. On the other hand, in the sporophytic SI S-glycoprotein gene is expressed in the papillar cells of the stigma as well as in e sporophytic tape is cells of anther wall. Recognition and rejection of self pollen in the gametophytic type is the reaction between the ribonuclease in the transmitting tissue of the style and the protein in the cytoplasm of pollen tube, whereas in the sporophytic system the inhibition of selfed pollen is caused by the interaction between the Sycoprotein in the wall of stigmatic papillar cell and the tapetum-origin protein deposited on the outer wall of the pollen grain. The claim that the S-allele-associated proteins are involved in recognition and rejection of self pollen has been made merely based on indirect evidence. Recently it has been verified that inhibition of synthesis of S$_3$ protein in Petunia inflata plants of S$_2$S$_3$ genotype by the antisense S$_3$ gene resulted in failure of the transgenic plant to reject S$_3$ pollen and that expression of the transgenic encoding S$_3$ protein in the S$_1$S$_2$ genotype confers on the transgenic plant the ability to reject S$_3$ pollen. These finding Provide direct evidence that S-proteins control the s elf-incompatibility behavior of the pistil.

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Surface glycoproteins determine the feature of the 2009 pandemic H1N1 virus

  • Kim, Jin Il;Lee, Ilseob;Park, Sehee;Park, Man-Seong
    • BMB Reports
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    • 제45권11호
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    • pp.653-658
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    • 2012
  • After the outbreak of the swine-origin influenza A H1N1 virus in April 2009, World Health Organization declared this novel H1N1 virus as the first pandemic influenza virus (2009 pH1N1) of the $21^{st}$ century. To elucidate the characteristics of 2009 pH1N1, the growth properties of A/Korea/01/09 (K/09) was analyzed in cells. Interestingly, the maximal titer of K/09 was higher than that of a seasonal H1N1 virus isolated in Korea 2008 (S/08) though the RNP complex of K/09 was less competent than that of S/08. In addition, the NS1 protein of K/09 was determined as a weak interferon antagonist as compared to that of S/08. Thus, in order to confine genetic determinants of K/09, activities of two major surface glycoproteins were analyzed. Interestingly, K/09 possesses highly reactive NA proteins and weak HA cell-binding avidity. These findings suggest that the surface glycoproteins might be a key factor in the features of 2009 pH1N1.

Protective Immunity Induced by Systemic and Mucosal Delivery of DNA Vaccine Expressing Glycoprotein B of Pseudorabies Virus

  • Yoon, Hyun-A;Han, Young-Woo;Aleyas, Abi George;George, June Abi;Kim, Seon-Ju;Kim, Hye-Kyung;Song, Hee-Jong;Cho, Jeong-Gon;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • 제18권3호
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    • pp.591-599
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    • 2008
  • A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.