A solid-phase lectin-binding assay for the characterization of glycan terminator on cell surface
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Keywords:
cancer biomarkers, sialyltransferases, glycan terminator, lectins, starvation.Abstract
Cell surface proteins in mammals are typically elaborated with a complex array of glycans. N-acetyl neuraminic acids (abbreviated as Sialic acids), are usually found at the non-reducing terminal position of these glycans. This terminal glycan sialylation imparts a negative charge at physiological pH values and mediates many biological functions. Here,we utilize two human mammary epithelial cell lines, MCF10A (breast normal cells) and MCF7 (breast cancer cells)as a model system to show differential glycan terminator when treated with sialic acid under nutrient deprivation.Under starved condition, sialic acid treatment of both cells resulted increased activities of α2→3/6 sialyltransferases as demonstrated by lectin solid phase assay. The presence of increased sialyltransferase expression is corroborated by stronger binding with sialic acid-specific lectins such as (Sambucus nigra agglutinin, SNA) and (Maackia amurensis agglutinin I, MAL-I). However, MAL-I binding discriminates malignant cells from normal cells suggesting a preferential increase of Neu5Acα2→3Gal on the SA-treated malignant cell surface.References
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2 Ohtsubo K. and Marth J.D. Glycosylation in cellular mechanisms of health and disease // Cell. 2006. V. 126. P. 855-67.
3 Exfoliation D. Turns that favor glycosylation // Science/ 2011. V. 331. P. 507.
4 Du J., Meledeo M.A., Wang Z., Khanna H.S., Paruchuri V.D.P. and Yarema K.J. Metabolic glycoengineering: Sialic acid and
beyond // Glycobiology. 2009. V. 12. P. 1382-1401.
5 Chaffer C.L. and Weinberg R.A. A perspective on cancer cell metastasis // Science. 2011. V. 331. P. 1559-64.
6 Sharon N. Lectins: Carbohydrate-specific Reagents and Biological Recognition Molecules // J. Biol. Chem. 2007. V. 282. P.
2753-2764.
7 Qian J., Zhu C.H., Tang S., Shen A.J., Ai J., Li J., Geng M.Y. and Ding J. α2,6-Hyposialylation of c-Met abolishes cell motility
of ST6Gal-I-knockdown HCT116 cells // Acta Pharm. Sinic. 2009. V. 30. P. 1039-1045.
8 Meany D.L. and Chan D.W. Aberrant glycosylation associated with enzymes as cancer biomarkers // Clin. Proteomics. 2011.
V. 8. P.1-14.
9 Picco G., Julien, S., Brockhausen I., Beatson R., Antonopoulos A., Haslam S., Mandel U., Dell A., Pinder S., Papadimitriou
J. and Burchell J. Over-expression of ST3Gal-I promotes mammary tumorigenesis // Glycobiology. 2010. V. 20 P. 1241-50.
10 Ohtsubo K., and Marth J.D. Glycosylation in cellular mechanisms of health and disease // Cell. 2006. V. 126. P. 855-67.
11 Harduin-Lepers A., Vallejo-Ruiz V., Krzewinski-Recchi M.A., Samyn-Petit B., Julien S. and Delannoy P. The human
sialyltransferase family // Biochimie. 2001. V. 83. P. 727-37.
12 Hakomori S. Glycosylation defining cancer malignancy: new wine in an old bottle // Proc. Natl. Acad. Sci. USA. 2002. V. 99.
P. 10231–33.
13 Agard N.J. and Bertozzi C.R. Chemical approaches to perturb, profile, and perceive glycans // Acc. Chem. Res. 2009. V. 42.
P. 788-97.
14 Mariño K., Bones J., Kattla J.J. and Rudd P.M. A systematic approach to protein glycosylation analysis: a path through the
maze // Nat. Chem. Biol. 2010. V. 6. P. 713-23.
15 Varki N.M. and Varki A. Diversity in cell surface sialic acid presentations: implications for biology and disease // Lab. Invest.
2007. V. 87. P. 851-57.
16 Dennis J.W., Nabi I.R. and Demetriou M. Metabolism, cell surface organization, and disease // Cell. 2009. V. 139. P. 1229-
1241.
17 Fuster M.M. and Esko J.D. The sweet and sour of cancer: glycans as novel therapeutic targets // Nat. Rev. Cancer. 2005. V. 5.
P. 526-542.
18 Geisler C. and Jarvis D.L. Letter to the Glyco-Forum: Effective glycoanalysis with Maackia amurensis lectins requires a clear
understanding of their binding specificities // Glycobiology. 2011. V. 21. P. 988-993.
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Badr, H. A., Djansugurova, L. B., & Ahmad, H. (2015). A solid-phase lectin-binding assay for the characterization of glycan terminator on cell surface. Experimental Biology, 57(1). Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/228
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HUMAN and ANIMAL PHYSIOLOGY
