Glycosylation and epitope mapping of the 5T4 glycoprotein oncofoetal antigen

The Biochemical Journal
David M ShawP L Stern

Abstract

The human 5T4 oncofoetal antigen is a focus for development of several antibody-directed therapies on the basis of the murine monoclonal antibody against 5T4 (mAb5T4), which recognizes a conformational epitope. 5T4 molecules are highly N-glycosylated transmembrane glycoproteins whose extracellular domain contains two regions of leucine-rich repeats (LRRs) and associated flanking regions, separated by an intervening hydrophilic sequence. Using a series of deletion and mutated cDNA constructs as well as chimaeras with the murine homologue, we have mapped the mAb5T4 epitope to the more membrane-proximal LRR2 or its flanking region. Analysis of the glycosylation of the seven consensus Asp-Xaa-Ser/Thr sites was consistent with all of the sites being glycosylated. A combination of two high-mannose chains (predominantly octasaccharide) and five mostly sialylated bi-, tri- and tetra-antennary complex chains with minor quantities of core fucose were detected. The two glycosylation sites, which are the most likely to have predominantly high-mannose chains, are in the only two regions that show significant differences between the human and the 81% identical mouse sequence. A site-directed mutation, which abolished glycosylation at one of ...Continue Reading

References

Jan 15, 1990·International Journal of Cancer. Journal International Du Cancer·N Hole, P L Stern
Sep 4, 1984·Journal of Immunological Methods·H Towbin, J Gordon
Jun 1, 1995·Current Opinion in Structural Biology·B Kobe, J Deisenhofer
Oct 1, 1994·Trends in Biochemical Sciences·B Kobe, J Deisenhofer
Sep 27, 1996·International Journal of Cancer. Journal International Du Cancer·C J CarsbergP L Stern
Jul 17, 1998·Methods in Molecular Biology·S Prime, T Merry
Jul 17, 1998·Methods in Molecular Biology·M J Davies, E F Hounsell
Oct 27, 1998·Trends in Biochemical Sciences·K Drickamer, M E Taylor
Dec 17, 1998·European Journal of Gastroenterology & Hepatology·T StarzynskaP L Stern
Jun 29, 2000·Molecular Biotechnology·N K WongE F Hounsell
Aug 10, 2000·Journal of Virology·A Mirazimi, L Svensson
Jan 19, 2002·Biochemical and Biophysical Research Communications·Abida AwanPeter L Stern

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Citations

Jul 9, 2004·Biochimica Et Biophysica Acta·Said DermimePeter L Stern
Sep 20, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Hui-Rong JiangPeter L Stern

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