Lectins as Tools to Select for Glycosylated Proteins
Glycosylation has been recognized as one of the most important modifications on proteins. The interactions between proteins and glycans are known to play an important role in many biological processes. Lectins are carbohydrate-binding proteins that can specifically interact with and select for carbohydrate structures. The technique of lectin affinity chromatography takes advantage of this specific interaction and enables the selection and purification of glycoproteins with carbohydrate structures complementary to the lectin-binding site. Depending on the carbohydrate specificity of the lectin glycoprotein fractions enriched for example, high mannose or complex N-glycans or O-glycans can be obtained. Afterward both the protein part and the glycan part can be analyzed in more detail allowing the identification of the interacting partners and the type of glycans involved.
Key wordsAffinity chromatography carbohydrate specificity glycan glycoprotein lectin
The financial support of the Research Council of Ghent University and Fund for Scientific Research-Flanders (G.0022.08) is gratefully acknowledged.
- 1.Van Damme, E. J. M., Peumans, W. J., Barre, A., and Rougé, P. (1998) Plant lectins: a composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles. Crit Rev Plant Sci 17, 575–692.Google Scholar
- 2.Van Damme, E. J. M., Peumans, W. J., Pusztai, A., and Bardocz, S. (1998) Handbook of Plant Lectins: Properties and Biomedical Applications, John Wiley & Sons, Chichester, p. 452.Google Scholar
- 5.Van Damme, E. J. M., Smith, D. F., Cummings, R., and Peumans, W. J. (2011) Glycan arrays to decipher the specificity of plant lectins. In: The Molecular Immunology of Complex Carbohydrates, A. M. Wu, ed., Kluwer Academic/Plenum Publishers, New York, 841–854.Google Scholar
- 13.Van Damme, E. J. M., Barre, A., Rougé, P., Van Leuven, F., and Peumans, W. J. (1996) The NeuAc (α-2,6)-Gal/GalNAc binding lectin from elderberry (Sambucus nigra) bark, a type 2 ribosome inactivating protein with an unusual specificity and structure. Eur J Biochem 235, 128–37.PubMedCrossRefGoogle Scholar
- 14.Wang, W.-C. and Cummings, R. D. (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 α-2,3 to penultimate galactose residues. J Biol Chem 261, 4576–85.Google Scholar
- 17.Bador, M., Cabrera, G., Stadlmann, J., Lerouge, P., Cremata, J. A., Gomord, V., et al. (2009) N-glycosylation of plant recombinant pharmaceuticals. In: Recombinant Proteins from Plants, Methods and Protocols, Springer Protocols, L. Faye and V. Gomord, eds., Humana Press, New York, pp. 239–64.Google Scholar
- 20.Lannoo, N., Peumans, W. J., Van Pamel, E., Alvarez, R., Xiong, T.-C., Hause, G., et al. (2006) Localization and in vitro binding studies suggest that the cytoplasmic/nuclear tobacco lectin can interact in situ with high-mannose and complex N-glycans. FEBS Lett 580, 6329–37.PubMedCrossRefGoogle Scholar