Molecular identification of lectin binding sites differentiating sialylation of epidermal growth factor receptor
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Keywords:
cancer biomarkers, epidermal growth factor receptor, , sialylation, lectins, starvationAbstract
The epidermal growth factor receptor (EGFR), family of receptor tyrosine kinases, also known as ErbB1 or HER1, plays crucial roles in the development of multicellular organisms. Mutations and over-expression of the EGF-receptors have been implicated in a variety of diseases. The extracellular, ligand binding regions of EGFR are quite heavily sialylated. Here we designed a lectin-metabolic assay that allows direct comparison of cell surface sialylation between health and disease states. We utilize two human mammary epithelial cell lines, HB4A (breast normal cells) and T47D (breast cancer cells) as a model system for the assessment of differential sialylation extracellular domain of ErbB1. Under starved condition, sialic acid treatment of both cells resulted increased lectin fluorescence signals that indicated the accumulation of more sialic acids in the metabolic state as demonstrated by Western blotting and immuno-precipitation. Furthermore, lectin-precipitation showed a very strong MAL-I (Maackia amurensis agglutinin I) binding on the EGFreceptors of sialic acid treated T47D cells suggesting an increase of Neu5Acα2→3Gal on the cell surface. The MAL-I lectin can be used for discrimination at molecular level between healthy and diseased cells.References
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3 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.
4 Sharon N. Lectins: Carbohydrate-specific Reagents and Biological Recognition Molecules // J. Biol. Chem. 2007. V. 282. P.
2753-64.
5 Varki N.M. and Varki A. Diversity in cell surface sialic acid presentations: implications for biology and disease // Lab. Invest.
2007. V. 87. P. 851-857.
6 Rudd P.M., Elliott T., Cresswell P., Wilson I.A. and Dwek R.A. Glycosylation and the immune system // Science. 2001. V. 291
P. 2370-6.
7 Wang X., Inoue S. and Gu J. Dysregulation of TGF-h1 receptor activation leads to abnormal lung development and
emphysema-like phenotype in core fucose-deficient mice // Proc Natl Acad Sci U S A. 2005. V. 102 P. 15791-6.
8 Tsuda T., Ikeda Y. and Taniguchi N. The Asn-420–linked sugar chain in human epidermal growth factor receptor suppresses
ligand-independent spontaneous oligomerization. Possible role of a specific sugar chain in controllable receptor activation // J. Biol.
Chem. 2000. V.275 P. 21988-94.
9 Looger L.L., Dwyer M.A., Smith J.J. and Hellinga H.W. Computational design of receptor and sensor proteins with novel
functions // Nature. 2003. V. 423 P. 185-90.
10 Hanash S.M., Pitteri S.J. and Faca V.M. Mining the plasma proteome for cancer biomarkers // Nature. 2009. V. 452 P. 571-79.
11 Dennis J.W., Nabi I.R. and Demetriou M. Metabolism, cell surface organization, and disease // Cell. 2009. V. 139. P. 1229-41.
12 Dube D.H. and Bertozzi C.R. Glycans in cancer and inflammation: potential for therapeutics and diagnostics // Nat. Rev. Drug
Discov. 2005. V. 4 P. 477-488.
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P. 526-42.
14 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., & Li, C.-Z. (2015). Molecular identification of lectin binding sites differentiating sialylation of epidermal growth factor receptor. Experimental Biology, 57(1), 170–174. Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/229
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HUMAN and ANIMAL PHYSIOLOGY
