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Use of Lectin for Detection of Agalactosyl IgG

Protocol
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 9)

Abstract

The immunoglobulin G (IgG) molecule contains two biantennary complextype oligosaccharide chains, each linked to the heavy chain at asparagine 297 within the CH2 domain (1) (see Note 1). X-ray crystallographic analysis suggested that the sugar chains of IgG play a role in maintaining the 3D structure of its Fc portion by bridging the two CH2 domains (2-4). Although these sugar chains can possess the complete structure shown in Fig. 1, normally only 25% of the sugar chains are sialylated, which is unusual because the sugar chains of other serum glycoproteins are highly sialylated Also characteristic is the extremely high microheterogeneity resulting from the presence or absence of the two galactose (Gal), the bisecting N-acetylglucosamine (GlcNAc), and the fucose (Fuc) residues (1).
Open image in new windowFig. 1.
Fig. 1.

Structure of asparagine-linked sugar chain of human IgG

Keywords

Juvenile Rheumatoid Arthritis Sugar Chain Laemmli Sample Buffer Galactose Content Lectin Binding Assay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. 1.
    Kobata, A (1990) Function and pathology of the sugar chains of human immunoglobulin G Glycobiology 1, 5–8PubMedCrossRefGoogle Scholar
  2. 2.
    Deisenhofer, J. (1981) Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from Staphylococcus aureus at 2 9-and 2.8-Å resolution. Biochemistry 20, 2361–2370PubMedCrossRefGoogle Scholar
  3. 3.
    Sutton, B J and Phillips, D. C (1983) The three-dimensional structure of the carbohydrate within the Fc fragment of immunoglobulin G Bcochem Soc Trans 11, 130–132Google Scholar
  4. 4.
    Parekh, R. B, Dwek, R A, Sutton, B J, Fernandes, D J, Leung, A, Stanworth, D., Rademacher, T. W, Mizuochi, T, Taniguchi, T., Matsuta, K, Takeuchi, F, Nagano, Y, Miyamoto, T, and Kobata, A (1985) Association of rheumatoid arthritis and primary osteoarthritis with changes in the glycosylation pattern of total serum IgG Nature 316, 452–457.PubMedCrossRefGoogle Scholar
  5. 5.
    Mullinax, F. (1975) Abnormality of IgG structure in rheumatoid arthritis and systemic lupus erythematosus. Arthritis Rheum. 18, 417Google Scholar
  6. 6.
    Parekh, R B., Roitt, I M, Isenberg, D A., Dwek, R. A., Anzell, B. M., and Rademacher, T W (1988) Galactosylation of IgG associated oligosaccharides reduction in patients with adult and juvenile onset rheumatoid arthritis and relation to disease activity Lancet ii, 964–969Google Scholar
  7. 7.
    Rook, G A. W, Steele, J., Brealey, R., Whyte, A, Isenberg, D., Sumar, N, Nelson, J. L, Bodman, K B, Young, A, Roitt, I. M, Williams, P, Scragg, I, Edge, C J, Arkwright, P D, Ashford, D., Wormald, M, Rudd, P, Redman, C W. G., Dwek, R A, and Rademacher, T W (1991) Changes in IgG glycoform levels are associated with remission of arthritis during pregnancy J Autoimmunity 4, 779–794.CrossRefGoogle Scholar
  8. 8.
    Tsuchiya, N., Endo, T, Matsuta, K, Yoshinoya, S, Takeuchi, F, Nagano, Y, Shiota, M, Furukawa, K, Kochibe, N, Ito, K, and Kobata, A (1993) Detection of glycosylation abnormality in rheumatoid IgG using N-acetylglucosamine-specific Psathyrella velutina lectin J Immunol. 151, 1137–1146PubMedGoogle Scholar
  9. 9.
    Parekh, R, Roitt, I, Isenberg, D, Dwek, R., and Rademacher, T (1988) Age-related galactosylation of the N-linked oligosaccharides of human serum IgG J Exp Med 167,1731–1736PubMedCrossRefGoogle Scholar
  10. 10.
    Tomana, M, Schrohenloher, R E, Koopman, W J, Alarcon, G S, and Paul, W A (1988) Abnormal glycosylation of serum IgG from patients with chronic inflammatory diseases Arthritis Rheum 31,333–338PubMedCrossRefGoogle Scholar
  11. 11.
    Dubé, R, Rook, G A W., Steele, J, Brealey, R, Dwek, R, Rademacher, T, and Lennard-Jones, J (1990) Agalactosyl IgG in inflammatory bowel disease correlation with C-reactive protein Gut 31, 431–434PubMedCrossRefGoogle Scholar
  12. 12.
    Bond, A, Cooke, A, and Hay, F C (1990) Glycosylation of IgG, immune complexes and IgG subclasses in the MRL-lpr/lpr mouse model of rheumatoid arthritis. Eur J Immunol 20, 2229–2233PubMedCrossRefGoogle Scholar
  13. 13.
    Mizuochi, T, Hamako, J, Nose, M, and Titani, K (1990) Structural changes in the oligosaccharide chains of IgG in autoimmune MRL/Mp-lpr/lpr mice. J Immunol. 145, 1794–1798PubMedGoogle Scholar
  14. 14.
    Thompson, S. J, Hitsumoto, Y, Zhang, Y W, Rook, G A W, and Elson, C J (1992) Agalactosyl IgG in pristane-induced arthritis. Pregnancy affects the incidence and severity of arthritis and the glycosylation status of IgG Clin Exp Immunol. 89,434–438.PubMedCrossRefGoogle Scholar
  15. 15.
    Furukawa, K. and Kobata, A. (1991) IgG galactosylation-its biological significance and pathology Mol Immunol 28, 1333–1340.PubMedCrossRefGoogle Scholar
  16. 16.
    Bodman, K. B, Sumar, N., MacKenzie, L E, Isenberg, D A, Hay, F C, Roitt, I M, and Lydyard, P. M (1992) Lymphocytes from patients with rheumatoid arthritis produce agalactosylated IgG in vitro. Clin Exp. Immunol 88, 420–423.CrossRefGoogle Scholar
  17. 17.
    Furukawa, K, Matsuta, K, Takeuchi, F, Kosuge, E, Miyamoto, T., and Kobata, A (1990) Kinetic study of a galactosyltransferase in the B cells of patients with rheumatoid arthritis Int. Immunol 2,105–112PubMedCrossRefGoogle Scholar
  18. 18.
    Hirano, T, Matsueda, T, Turner, M, Miyasaka, N, Buchan, G, Tang, B, Sato, K, Shimizu, M., Maini, R., Feldmann, M, and Kishimoto, T. (1988) Excessive production of interleukin 6/B cell stimulatory factor-2 in rheumatoid arthritis. Eur J Immunol. 18, 1797–1801PubMedCrossRefGoogle Scholar
  19. 19.
    Nakao, H, Nishikawa, A., Nishiura, T, Kanayama, Y., Tarui, S., and Taniguchi, N. (1991) Hypogalactosylation of immunoglobulin G sugar chains and elevated serum interleukin 6 in Castleman’s disease. Clin Chim Acta 197, 221–228PubMedCrossRefGoogle Scholar
  20. 20.
    Hitsumoto, Y, Thompson, S J., Zhang, Y W., Rook, G A W, and Elson, C J (1992) Relationship between interleukin 6, agalactosyl IgG and pristane-induced arthritis Autoimmunity 11, 247–254PubMedCrossRefGoogle Scholar
  21. 21.
    Axford, J. S, Sumar, N, Alavi, A, Isenberg, D A., Young, A, Bodman, K B, and Roitt, I M (1992) Changes in normal glycosylation mechanisms in autoimmune rheumatic disease J Clin Invest. 89, 1021–1031PubMedCrossRefGoogle Scholar
  22. 22.
    Nose, M and Wigzell, H. (1983) Biological significance of carbohydrate chains on monoclonal antibodies Proc Natl Acad Sci USA 80, 6632–6636PubMedCrossRefGoogle Scholar
  23. 23.
    Leatherbarrow, R J, Rademacher, T W, Dwek, R A, Woof, J M, Clark, A, and Burton, D. (1985) Effecter functions of a monoclonal aglycosylated mouse IgG2a binding and activation of complement component Cl and interaction with human monocyte Fc receptor. Mol Immunol 22,407–415PubMedCrossRefGoogle Scholar
  24. 24.
    Tsuchiya, N, Endo, T, Matsuta, K, Yoshinoya, S, Aikawa, T, Kosuge, E, Takeuchi, F., Miyamoto, T., and Kobata, A. (1989) Effects of galactose depletion from oligosaccharide chanis on immunological activities of human IgG J Rheumatol 16,285–290PubMedGoogle Scholar
  25. 25.
    Newkirk, M M, Lemmo, A, and Rauch, J. (1990) Importance of the IgG isotype, not the state of glycosylation, in determining human rheumatoid factor binding Arthritis Rheum 33, 800–809PubMedCrossRefGoogle Scholar
  26. 26.
    Lund, J., Takahashi, N, Pound, J D, Goodall, M, Nakagawa, H, and Jefferis, R (1995) Oligosaccharide-protein interaction in IgG can modulate recognition by Fcγ receptors FASEB J 9, 115–119PubMedGoogle Scholar
  27. 27.
    Young, A, Sumar, N., Bodman, K, Goyal, S, Sinclair, H Roitt, I, and Isenberg, D (1991) Agalactosyl IgG an aid to differential diagnosis in early synovitis Arthritis Rheum 34, 1425–1429PubMedCrossRefGoogle Scholar
  28. 28.
    Mizuochi, T, Taniguchi, T, Shimizu, A, and Kobata, A (1982) Structural and numerical variations of the carbohydrate moiety of immunoglobulin G J Immunol 129,2016–2019PubMedGoogle Scholar
  29. 29.
    Sumar, N., Bodman, K B., Rademacher, T. W, Dwek, R. A., Williams, P, Parekh, R. B., Edge, J., Rook, G A W, Isenberg, D. A., Hay, F C, and Roitt, I M (1990) Analysis of glycosylation changes in IgG using lectins. J Immunol Methods 131,127–136PubMedCrossRefGoogle Scholar
  30. 30.
    Parkkinen, J. (1989) Aberrant lectin-binding activity of immunoglobulin G in serum from rheumatoid arthritis patients. Clin Chem 35,1638–1643PubMedGoogle Scholar
  31. 31.
    Kochibe, N. and Matta, K L (1989) Purification and properties of an N-acetylglucosamine-specific lectin from Psathyrella velutina mushroom J Biol Chem 264, 173–177PubMedGoogle Scholar
  32. 32.
    Endo, T, Ohbayashi, H, Kanazawa, K, Kochibe, N, and Kobata, A (1992) Carbohydrate binding specificity of immobilized Psathyrella velutina lectin J Biol Chem 267,707–713PubMedGoogle Scholar
  33. 33.
    Sasso, E. H., Silverman, G J., and Mannik, M. (1989) Human IgM molecules that bind staphylococcal protein A contain VHIII H chains J Immunol 142,2778–2783PubMedGoogle Scholar
  34. 34.
    Sasso, E. H, Silverman, G J, and Mannik, M (1991) Human IgA and IgG F(ab’)2 that bind to staphylococcal protein A belong to the VHIII subgroup. J Immunol 147,1877–1883PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  1. 1.Department of Internal Medicine and Physical Therapy, Faculty of MedicineUniversity of TokyoTokyoJapan
  2. 2.Department of GlycobiologyTokyo Metropolitan Institute of GerontologyTokyoJapan
  3. 3.Department of Biology, Faculty of EducationGunma UniversityGunmaJapan
  4. 4.Department of Internal Medicine and Physical Therapy, Faculty of MedicineUniversity of TokyoTokyoJapan
  5. 5.Tokyo Metropolitan Institute of GerontologyTokyoJapan

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