Glycan Markers of Human Stem Cells Reassigned with Beam Search Arrays

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Authors
Wu, N.
Silva, L.M.
Liu, Y.
Zhang, Y.
Gao, C.
Zhang, F.
Fu, L.
Peng, Y.
Linhardt, Robert J.
Kawasaki, T.
Issue Date
2019
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Abstract
Glycan antigens recognized by monoclonal antibodies have served as stem cell markers. To understand regulation of their biosynthesis and their roles in stem cell behavior precise assignments are required. We have applied state-of-the-art glycan array technologies to compare the glycans bound by five antibodies that recognize carbohydrates on human stem cells. These are: FC10.2, TRA-1-60, TRA-1-81, anti-i and R-10G. Microarray analyses with a panel of sequence-defined glycans corroborate that FC10.2, TRA-1-60, TRA-1-81 recognize the type 1-(Galβ-3GlcNAc)-terminating backbone sequence, Galβ-3GlcNAcβ-3Galβ-4GlcNAcβ-3Galβ-4GlcNAc, and anti-i, the type 2-(Galβ-4GlcNAc) analog, Galβ-4GlcNAcβ-3Galβ-4GlcNAcβ-3Galβ-4GlcNAc, and we determine substituents they can accommodate. They differ from R-10G, which requires sulfate. By Beam Search approach, starting with an antigen-positive keratan sulfate polysaccharide, followed by targeted iterative microarray analyses of glycan populations released with keratanases and mass spectrometric monitoring, R-10G is assigned as a mono-sulfated type 2 chain with 6-sulfation at the penultimate N-acetylglucosamine, Galβ-4GlcNAc(6S)β-3Galβ-4GlcNAcβ-3Galβ-4GlcNAc. Microarray analyses using newly synthesized glycans corroborate the assignment of this unique determinant raising questions regarding involvement as a ligand in the stem cell niche.
Description
Molecular and Cellular Proteomics, 18, 1981–2002
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Full Citation
Glycan Markers of Human Stem Cells Reassigned with Beam Search Arrays, N. Wu, L. M. Silva, Y. Liu, Y. Zhang, C. Gao, F. Zhang, L. Fu, Y. Peng, R. J. Linhardt, T. Kawasaki, B. Mulloy, W. Chai, T. Feizi, Molecular and Cellular Proteomics, 18, 1981–2002, 2019.
Publisher
The American Society for Biochemistry and Molecular Biology, Inc.
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