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Preparation of Glycan Arrays Using Pyridylaminated Glycans

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Part of the Methods in Molecular Biology book series (MIMB,volume 1368)

Abstract

We describe the method to prepare neoglycoproteins from the conjugation of bovine serum albumin and pyridylaminated glycans. Large quantities of glycans (>1 mg) can be pyridylaminated and then converted to their 1-amino-1-deoxy derivatives by reaction with hydrogen followed by hydrazine. These pyridylaminated glycans can then be conjugated to bovine serum albumin via esterification with N-( m-maleimidobenzoyloxy)succinimide to form a neoglycoprotein, e.g., glycosylated bovine serum albumin. As a demonstration, we prepared High-mannose bovine serum albumin, which was immobilized on an activated glass slide. Then, we showed that the neoglycoprotein bind to Cy3-labeled Lens culinaris agglutinin, a mannose-specific plant lectin, as detected using an evanescent-field-activated fluorescence scanner system.

Key words

  • Pyridylamination
  • 1-Amino-1-deoxy derivative
  • Neoglycoprotein
  • Glycan array
  • Evanescent-field-activated fluorescence scanner

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References

  1. Trevino V, Falciani F, Barrera-Saldana HA (2007) DNA microarrays. A powerful genomic tool for biomedical and clinical research. Mol Med 13:527–541

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

  2. Kricka LJ, Master SR, Joos TO, Fortina P (2006) Current perspectives in protein array technology. Ann Clin Biochem 43:457–467

    CAS  CrossRef  PubMed  Google Scholar 

  3. Smith DF, Song X, Cummings RD (2010) Use of glycan microarrays to explore specificity of glycan-binding proteins. Methods Enzymol 480:417–444

    CAS  CrossRef  PubMed  Google Scholar 

  4. Rillahan CD, Paulson JC (2011) Glycan microarrays for decoding the glycome. Annu Rev Biochem 80:797–823

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

  5. Oyeleran O, Gildesleeve JC (2009) Glycan array. Recent advances and future challenges. Curr Opin Chem Biol 13:406–413

    CrossRef  Google Scholar 

  6. Fukui S, Feizi T, Galustian C, Lawson AM, Chai W (2002) Oligoglycan microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interaction. Nat Biotechnol 20:1011–1017

    CAS  CrossRef  PubMed  Google Scholar 

  7. Wang D, Liu S, Trummer BJ, Deng C, Wang A (2002) Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microtubes and host cells. Nat Biotechnol 20:275–281

    CAS  CrossRef  PubMed  Google Scholar 

  8. Robinson MJ, Sancho D, Slack EC, LeibundGut-Landmann S, Reis e Sousa C (2006) Myeloid C-type lectins in innate immunity. Nat Immunol 7:1258–1265

    CAS  CrossRef  PubMed  Google Scholar 

  9. Crocker PR, Paulson JC, Valki A (2007) Siglecs and their roles in the immune system. Nat Rev Immunol 7:255–266

    CAS  CrossRef  PubMed  Google Scholar 

  10. van Vliet SJ, den Dunnen J, Gringhuis SI, Geijtenbeek TB, van Kooyk Y (2007) Innate signaling and regulation of dendric cell immunity. Curr Opin Immunol 19:189–194

    Google Scholar 

  11. Blixt O, Head S, Mondala T, Scanlan C, Huflejt ME, Alvarez R, Bryan MC, Fazio F, Calarese D, Stevens J, Razi N, Stevens DJ, Skehel JJ, van Die I, Burton DR, Wilson IA, Cummings R, Bovin N, Wong CH, Paulson JC (2004) Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc Natl Acad Sci U S A 101:17033–17038

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

  12. Tateno H, Mori A, Uchiyama N, Yabe R, Iwaki J, Shikanai T, Angata T, Narimatsu H, Hirabayashi J (2008) Glycoconjugate microarray based on an evanescent-field fluorescence-assisted detection principle for investigation of glycan-binding proteins. Glycobiology 18:789–798

    CAS  CrossRef  PubMed  Google Scholar 

  13. Padler-Karavani V, Song X, Yu H, Hurtado-Ziola N, Huang S, Muthana S, Chokhawala HA, Cheng J, Verhagen A, Langereis MA, Kleene R, Schachner M, de Groot RJ, Lasanajak Y, Matsuda H, Schwab R, Chen X, Smith DF, Cummings RD, Varki A (2012) Cross-comparison of protein recognition of sialic acid diversity on two novel sialoglycan microarrays. J Biol Chem 287:22593–22608

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

  14. Hase S, Ikenaka T, Matsushima Y (1978) Structure analyses of oligoglycans by tagging of the reducing end sugars with a fluorescent compound. Biochem Biophys Res Commun 85:257–263

    CAS  CrossRef  PubMed  Google Scholar 

  15. Sumiyoshi W, Nakakita S, Miyanishi N, Hirabayashi J (2009) Strategic glycan elution map for the production of human-type N-linked oligoglycans: the case of hen egg yolk and white. Biosci Biotechnol Biochem 73:543–551

    CAS  CrossRef  PubMed  Google Scholar 

  16. Yashizawa Z, Sato T, Schmid K (1966) Hydrazinolysis of alpha-1-acid glycoprotein. Biochim Biophys Acta 121:417–420

    CrossRef  Google Scholar 

  17. Nakakita S, Sumiyoshi W, Miyanishi N, Hirabayashi J (2007) A practical approach to N-glycan production by hydrazinolysis using hydrazine monohydrate. Biochem Biophys Res Commun 362:639–645

    CAS  CrossRef  PubMed  Google Scholar 

  18. Hase S (1992) Conversion of pyridylamino sugar chains to 1-amino-1-deoxy derivatives, intermediates for tagging with fluorescein and biotin. J Biochem 112:266–268

    CAS  PubMed  Google Scholar 

  19. Kuno A, Uchiyama N, Koseki-Kuno S, Ebe Y, Takashima S, Yamada M, Hirabayashi J (2005) Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling. Nat Methods 2:851–856

    CAS  CrossRef  PubMed  Google Scholar 

  20. Lobl TJ, Mitchell MA, Maggiora LL (1990) SV40 large T-antigen nuclear signal analogues: successful nuclear targeting with bovine serum albumin but not low molecular weight fluorescent conjugates. Biopolymers 29:197–203

    CAS  CrossRef  PubMed  Google Scholar 

  21. Takahashi T, Kawakami T, Mizuno T, Minami A, Uchida Y, Saito T, Matsui S, Ogata M, Usui T, Sriwilaijaroen N, Hiramatsu H, Suzuki Y, Suzuki T (2013) Sensitive and direct detection of receptor binding specificity of highly pathogenic avian influenza A virus in clinical samples. PLoS One 8:e78125

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

  22. Xu R, de Vries RP, Zhu X, Nycholat CM, McBride R, Yu W, Paulson JC, Wilson IA (2013) Preferential recognition of avian-like receptors in human influenza A H7N9 viruses. Science 342:1230–1235

    PubMed Central  CAS  CrossRef  PubMed  Google Scholar 

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Acknowledgement

We are grateful to Professor Yasuo Suzuki (Chubu University, Japan) for providing the H1N1 influenza virus sample

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Correspondence to Shin-ichi Nakakita .

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Nakakita, Si., Hirabayashi, J. (2016). Preparation of Glycan Arrays Using Pyridylaminated Glycans. In: Li, P., Sedighi, A., Wang, L. (eds) Microarray Technology. Methods in Molecular Biology, vol 1368. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3136-1_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3136-1_16

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3135-4

  • Online ISBN: 978-1-4939-3136-1

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