A Human Pluripotent Stem Cell Surface N-Glycoproteome Resource Reveals Markers, Extracellular Epitopes, and Drug Targets

• Published in: Stem cell reports 2014-07, Vol.3 (1), p.185-203
• Main Authors: Boheler, Kenneth R., Bhattacharya, Subarna, Kropp, Erin M., Chuppa, Sandra, Riordon, Daniel R., Bausch-Fluck, Damaris, Burridge, Paul W., Wu, Joseph C., Wersto, Robert P., Chan, Godfrey Chi Fung, Rao, Sridhar, Wollscheid, Bernd, Gundry, Rebekah L.
• Format: Article
• Language: English
• Subjects: Cells, Cultured; Epitopes - analysis; Epitopes - immunology; Flow Cytometry; Glycoproteins - immunology; Glycoproteins - metabolism; Humans; Immunohistochemistry; Pluripotent Stem Cells - immunology; Pluripotent Stem Cells - metabolism; Proteome - analysis; Real-Time Polymerase Chain Reaction
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2014.05.002

Detailed knowledge of cell-surface proteins for isolating well-defined populations of human pluripotent stem cells (hPSCs) would significantly enhance their characterization and translational potential. Through a chemoproteomic approach, we developed a cell-surface proteome inventory containing 496 N-linked glycoproteins on human embryonic (hESCs) and induced PSCs (hiPSCs). Against a backdrop of human fibroblasts and 50 other cell types, >100 surface proteins of interest for hPSCs were revealed. The >30 positive and negative markers verified here by orthogonal approaches provide experimental justification for the rational selection of pluripotency and lineage markers, epitopes for cell isolation, and reagents for the characterization of putative hiPSC lines. Comparative differences between the chemoproteomic-defined surfaceome and the transcriptome-predicted surfaceome directly led to the discovery that STF-31, a reported GLUT-1 inhibitor, is toxic to hPSCs and efficient for selective elimination of hPSCs from mixed cultures. [Display omitted] •496 cell surface N-glycoproteins on hPSCs•N-glycosylation site identification dictates accessible epitopes•>30 positive and negative selection markers for hPSCs are validated•STF-31 is selectively toxic to hPSCs Gundry, Boheler, and colleagues reveal a cell surfaceome containing 496 N-linked glycoproteins identified by mass spectrometry on human pluripotent stem cells. More than 30 positive and negative markers were verified by orthogonal approaches. This resource led to the discovery that a small molecule, STF-31, is efficient for the selective elimination of pluripotent cells from mixed cultures.