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Determination of Glycan Motifs Using Serial Lectin Affinity Chromatography

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Lectins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1200))

Abstract

Serial lectin affinity chromatography is a convenient technique for characterizing glycan motifs (terminal glycan structures) of glycoproteins or released glycans. When these glycoconjugates are applied serially or in parallel to lectin-immobilized columns, information regarding the glycan motifs can be obtained. We demonstrate lectin affinity chromatographic methods for determining O-linked glycan structures of MUC1 purified from a breast cancer cell line, YMB-S, N-linked glycan structures of serum prostate-specific antigen from prostate cancer, and serum alkaline phosphatases from choriocarcinoma. These lectin-fractionated samples are analyzed quantitatively by measuring radioactivity, antigen contents are analyzed using enzyme-linked immunosorbent assay, and enzymatic activities are assessed.

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References

  1. Valki A (1993) Biological roles of oligosaccharides: all of theories are correct. Glycobiology 3:97–130

    Article  Google Scholar 

  2. Vliegenthart JFG, Kamerling JP (2007) 1H-NMR structural-reporter-group concepts in carbohydrate analysis. In: Kamerling JP (ed) Comprehensive glycoscience, vol 2. Elsevier Ltd, Oxford, pp 133–191

    Chapter  Google Scholar 

  3. Dell A, Morris HR (2001) Glycoprotein structure determination by mass spectrometry. Science 291:2351–2356

    Article  PubMed  CAS  Google Scholar 

  4. Geyer R, Geyer H (1994) Saccharide linkage analysis using methylation and other techniques. In: Lennarz WJ, Hart GW (eds) Methods in enzymology, vol 230. Academic, San Diego, pp 86–108

    Google Scholar 

  5. Cummings RD, Etzler ME (2009) Antibodies and lectins in glycan analysis. In: Varki A, Cummings RD, Esko JD, Freeze H, Hart G, Marth J (eds) Essentials of glycobiology, 2nd edn. Cold Spring Harbor Laboratory Press, New York, Chapter 45

    Google Scholar 

  6. Kobata A, Yamashita K (1993) Fractionation of oligosaccharides by serial affinity chromatography with use of immobilized lectin columns. In: Fukuda M, Kobata A (eds) Practical approach-glycoprotein analysis. Oxford University Press, New York, pp 103–125

    Google Scholar 

  7. Seko A, Ohkura T, Ideo H et al (2012) Novel O-linked glycans containing 6’-sulfo-Gal/GalNAc of MUC1 secreted from human breast cancer YMB-S cells: possible carbohydrate epitopes of KL-6(MUC1) monoclonal antibody. Glycobiology 22:181–195

    Article  PubMed  CAS  Google Scholar 

  8. Fukushima K, Satoh T, Baba S et al (2010) α-1,2-Fucosylated and β-N-acetylgalactosaminylated prostate-specific antigen as an efficient marker of prostatic cancer. Glycobiology 20:452–460

    Article  PubMed  CAS  Google Scholar 

  9. Fukushima K, Hara-Kuge S, Seko A et al (1998) Elevation of α2-6sialyltransferase and α1-2fucosyltransferase in human choriocarcinoma. Cancer Res 58:4301–4306

    PubMed  CAS  Google Scholar 

  10. Cuatrecasas P, Anfinsen CB (1971) Affinity chromatography. In: Jakohy WS (ed) Methods in enzymology. Academic, New York, pp 345–378

    Google Scholar 

  11. Ohkura T, Seko A, Hara-Kuge S et al (2002) Occurrence of secretory glycoprotein-specific GalNAcβ1-4GlcNAc sequence in N-glycans in MDCK cells. J Biochem 132:891–901

    Article  PubMed  CAS  Google Scholar 

  12. Ohkura T, Hada T, Higashino K et al (1994) Increase of fucosylated serum cholinesterase in relation to high risk groups for hepatocellular carcinomas. Cancer Res 54:55–61

    PubMed  CAS  Google Scholar 

  13. Yamashita K, Umetsu K, Suzuki T et al (1988) Carbohydrate binding specificity of immobilized Allomyrina dichotoma lectin II. J Biol Chem 263:17482–17489

    PubMed  CAS  Google Scholar 

  14. Yamashita K, Umetsu K, Suzuki T et al (1992) Purification and characterization of a Neu5Acα2 → 6Galβ1 → 4GlcNAc and HSO3 − → 6Galβ1 → 4GlcNAc specific lectin in tuberous roots of Trichosanthes japonica. Biochemistry 31:11647–11650

    Article  PubMed  CAS  Google Scholar 

  15. Wang WC, Cummings RD (1988) The immobilized leukoagglutinin from the seeds of Maackia amurensis binds with high affinity to complex-type Asn-linked oligosaccharides containing terminal sialic acid-linked α2,3 to penultimate galactose residues. J Biol Chem 263:4576–4585

    PubMed  CAS  Google Scholar 

  16. Shibuya N, Goldstein IJ, Broekaert WF et al (1987) The elderberry (Sambucus nigra L.) bark lectin recognizes the Neu5Ac(α2-6)Gal/GalNAc sequence. J Biol Chem 262:1596–1601

    PubMed  CAS  Google Scholar 

  17. Kochibe N, Matta KL (1989) Purification and properties of an N-acetylglucosamine-specific lectin from Psathyrella velutina mushroom. J Biol Chem 264:173–177

    PubMed  CAS  Google Scholar 

  18. Ideo H, Seko A, Ohkura T et al (2002) High-affinity binding of recombinant human galectin-4 to SO3 − → 3Galβ1 → 3GalNAc pyranoside. Glycobiology 12:199–208

    Article  PubMed  CAS  Google Scholar 

  19. Smith DF, Torres BV (1989) Lectin affinity chromatography of glycolipids and glycolipid-derived oligosaccharides. In: Ginsburg V (ed) Methods in enzymology, vol 179. Academic, New York, pp 30–45

    Google Scholar 

  20. Hindgaul O, Norberg T, Le Pendu J et al (1982) Synthesis of type 2 human blood-group antigenic determinants. The H, X and Y haptens and variations of the H type 2 determinants probes for the combining site of the lectin I of Ulex europaeus. Carbohydr Res 109:109–142

    Article  Google Scholar 

  21. Kornfeld K, Reitman ML, Kornfeld R (1981) The carbohydrate-binding specificity of pea and lentil lectins: fucose is an important determinant. J Biol Chem 256:6633–6640

    PubMed  CAS  Google Scholar 

  22. Yamashita K, Kochibe N, Ohkura T et al (1985) Fractionation of L-fucose-containing oligosaccharides on immobilized Aleuria aurantia lectin. J Biol Chem 260:4688–4693

    CAS  Google Scholar 

  23. Yamashita K, Totani K, Ohkura T et al (1987) Carbohydrate binding properties of complex-type oligosaccharides on immobilized Datura stramonium lectin. J Biol Chem 262:1602–1607

    PubMed  CAS  Google Scholar 

  24. Baenziger JU, Fiete D (1979) Structural determinants of concanavalin A specificity for oligosaccharides. J Biol Chem 254:2400–2407

    PubMed  CAS  Google Scholar 

  25. Hoyer LL, Roggentin P, Schauer R et al (1991) Purification and properties of cloned Salmonella typhimurium LT2 sialidase with virus: typical kinetic preference for sialyl α2-3 linkages. J Biochem 110:29–41

    Google Scholar 

  26. Uchida Y, Tsukada Y, Sugimori T (1974) Production of microbial neuraminidase induced by colominic acid. Biochim Biophys Acta 350:425–431

    Article  PubMed  CAS  Google Scholar 

  27. Fukushima K, Hada T, Higashino K et al (1998) Elevated serum levels of Trichosanthes japonica agglutinin-I binding alkaline phosphatase in relation to high-risk groups for hepatocellular carcinomas. Clin Cancer Res 4:2771–2777

    PubMed  CAS  Google Scholar 

  28. Kiyohara T, Terao T, Shioiri-Nakano K, Osawa T (1976) Purification and characterization of β-N-acetylhexosaminidase, and β-galactosidase from Streptococcus 6646K. J Biochem 80: 9–17

    PubMed  CAS  Google Scholar 

  29. Li YT, Li SC (1972) α-Mannosidase, β-N-acetylhexosaminidase, and β-galactosidase from jack bean meal. In: Ginsburg V (ed) Methods in enzymology. Academic, New York, pp 702–713

    Google Scholar 

  30. Endo T, Ohbayashi H, Ikehara Y et al (1988) Structural study on the carbohydrate moiety of human placental alkaline phosphatase. J Biochem 103:182–187

    PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by JSPS KAKENHI, Grant Number 24590345, in Japan.

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Authors and Affiliations

  1. Department of Histology and Cell Biology, School of Medicine, Yokohama City University, 3-9, Fukuura, Kanazawa-ku, 236-0004, Yokohama, Japan

    Katsuko Yamashita

  2. Department of Reproductive Biology, National Center for Child Health and Development, Tokyo, Japan

    Takashi Ohkura

Authors
  1. Katsuko Yamashita
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  2. Takashi Ohkura
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Correspondence to Katsuko Yamashita .

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Editors and Affiliations

  1. National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan

    Jun Hirabayashi

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Yamashita, K., Ohkura, T. (2014). Determination of Glycan Motifs Using Serial Lectin Affinity Chromatography. In: Hirabayashi, J. (eds) Lectins. Methods in Molecular Biology, vol 1200. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1292-6_7

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  • DOI: https://doi.org/10.1007/978-1-4939-1292-6_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1291-9

  • Online ISBN: 978-1-4939-1292-6

  • eBook Packages: Springer Protocols

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