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Mapping Specific Cellular Sialoglycans Using Glycosyltransferases
Author(s) -
Wu Zhengliang L.,
Person Anthony D.,
Burroughs Barbara,
Singh Ravinder,
Tatge Timothy,
Manning Timothy,
Wu Guoping,
Sackstein Robert,
Kalabokis Vassili
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.673.8
Subject(s) - glycan , sialic acid , neuraminic acid , chemistry , hela , mannose , biochemistry , epitope , mannose receptor , selectin , receptor , glycobiology , glycosylation , glycosyltransferase , antigen , biology , glycoprotein , cell , microbiology and biotechnology , cell adhesion molecule , immunology , in vitro , gene , macrophage
The spatial and temporal decoration of glycans with neuraminic acid (sialylation) on cell surface involves in many biological processes, ranging from providing a physical barrier, shielding galactose and mannose residues from respective receptors, to generating specific epitopes that guide cell adhesion events. For examples, sialylated Lewis X structure serves as a ligand for selectins and is essential for leukocyte homing process; sialylated T/Tn antigens are involved in oncogenic processes and cancer invasiveness; while α‐(2,3)‐linked sialic acids serve as receptors for avian influenza vial infection, α‐(2,6)‐linked sialic acids serve as receptors for human influenza viral infection. Mapping the distribution of these glycans on cell surface is essential for us to understand their biological roles. Here, using HELA, HUVEC and CHO cells as models, we describe techniques for imaging some specific sialo‐glycans using glycosyltransferases including ST6Gal1, ST3Gal1, ST6GalNAc1, FUT2, and FUT9, with respective clickable nucleotide sugar donors. Our data suggest that while small percentages of terminal lactosamine (precursors for Lewis X structure) and Tn antigen may not be sialylated, almost all N‐glycans and Core‐1 O‐glycans on HELA cells are sialylated. Our data also indicate that while N‐glycan and lactosamine are more enriched on cell edges, where they may participate in cell‐cell interactions, sialylated Core‐1 O‐glycan and Tn antigen are more evenly distributed on cell surface. Despite the notion that CHO cells show no evidence for ST6Gal1 gene expression, we found that CHO cells are covered with α‐(2,6)‐linked sialic acid, which is also confirmed by sialic acid probing on CHO expressed proteins. Support or Funding Information This work was supported by R&D Systems/Bio‐techne This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .