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Core saccharide dependence of sialyl Lewis X biosynthesis

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Abstract

The sialyl-Lewis X (SLex) determinant is important in leukocyte extravasation, metastasis and bacterial adhesion. The role of the protein, N-glycan and O-glycan core structures for the biosynthesis of SLex in vivo by fucosyltransferases (FucTs) is not known. Immunoglobulin G (IgG) Fc fusion proteins of α1-acid glycoprotein (AGP), P-selectin glycoprotein ligand-1 (PSGL-1) or CD43 were used to probe the specificity of FucT-III-VII expressed alone in 293T and COS cells or together with O-glycan core enzymes in Chinese hamster ovary (CHO)-K1 cells. Western blotting with the monoclonal antibodies CSLEX and KM93 showed that FucT-III and V-VII produced SLex on core 2 in CHO cells. Only FucT-V, -VI and, with low activity, -VII worked on core 3 on CD43/IgG, but no SLex was detected with CSLEX on PSGL-1/IgG with core 3. KM93 stained SLex on core 2, but was not reactive with SLex on core 3. FucT-III, V-VII made SLex on N-glycans of AGP/IgG in CHO, but not in COS and 293T cells, even though the same FucTs could make SLex on CD43/IgG and PSGL-1/IgG in these cells. Our results define the specificities of FucT-III-VII in SLex biosynthesis on O-glycans with different core structures and the fine specificity of the widely used anti-SLex monoclonal antibody, KM93.

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Acknowledgements

We thank Dr Brian Seed for kindly sharing expression plasmids.

This work was supported by the Swedish Research Council, grants K2005-06X-13031-07A and K2005-06BI-15356-01A. J.H. and J.L. were both supported by the program “Glycoconjugates in Biological Systems” financed by the Swedish Foundation for Strategic Research.

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  1. Division of Clinical Immunology and Transfusion Medicine, F79, Karolinska Institute, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden

    Jonas Löfling & Jan Holgersson

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Löfling, J., Holgersson, J. Core saccharide dependence of sialyl Lewis X biosynthesis. Glycoconj J 26, 33–40 (2009). https://doi.org/10.1007/s10719-008-9159-z

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