TY - JOUR
T1 - Precision mapping of the human O-GalNAc glycoproteome through SimpleCell technology
AU - Steentoft, Catharina
AU - Vakhrushev, Sergey Y
AU - Joshi, Hiren J
AU - Kong, Yun
AU - Vester-Christensen, Malene B
AU - Schjoldager, Katrine Ter-Borch Gram
AU - Lavrsen, Kirstine
AU - Dabelsteen, Sally
AU - Pedersen, Nis Borbye
AU - da Silva, Lara Patricia Marcos
AU - Gupta, Ramneek
AU - Paul Bennett, Eric
AU - Mandel, Ulla
AU - Brunak, Søren
AU - Wandall, Hans H
AU - Levery, Steven B
AU - Clausen, Henrik
PY - 2013/5/15
Y1 - 2013/5/15
N2 - Glycosylation is the most abundant and diverse posttranslational modification of proteins. While several types of glycosylation can be predicted by the protein sequence context, and substantial knowledge of these glycoproteomes is available, our knowledge of the GalNAc-type O-glycosylation is highly limited. This type of glycosylation is unique in being regulated by 20 polypeptide GalNAc-transferases attaching the initiating GalNAc monosaccharides to Ser and Thr (and likely some Tyr) residues. We have developed a genetic engineering approach using human cell lines to simplify O-glycosylation (SimpleCells) that enables proteome-wide discovery of O-glycan sites using 'bottom-up' ETD-based mass spectrometric analysis. We implemented this on 12 human cell lines from different organs, and present a first map of the human O-glycoproteome with almost 3000 glycosites in over 600 O-glycoproteins as well as an improved NetOGlyc4.0 model for prediction of O-glycosylation. The finding of unique subsets of O-glycoproteins in each cell line provides evidence that the O-glycoproteome is differentially regulated and dynamic. The greatly expanded view of the O-glycoproteome should facilitate the exploration of how site-specific O-glycosylation regulates protein function.
AB - Glycosylation is the most abundant and diverse posttranslational modification of proteins. While several types of glycosylation can be predicted by the protein sequence context, and substantial knowledge of these glycoproteomes is available, our knowledge of the GalNAc-type O-glycosylation is highly limited. This type of glycosylation is unique in being regulated by 20 polypeptide GalNAc-transferases attaching the initiating GalNAc monosaccharides to Ser and Thr (and likely some Tyr) residues. We have developed a genetic engineering approach using human cell lines to simplify O-glycosylation (SimpleCells) that enables proteome-wide discovery of O-glycan sites using 'bottom-up' ETD-based mass spectrometric analysis. We implemented this on 12 human cell lines from different organs, and present a first map of the human O-glycoproteome with almost 3000 glycosites in over 600 O-glycoproteins as well as an improved NetOGlyc4.0 model for prediction of O-glycosylation. The finding of unique subsets of O-glycoproteins in each cell line provides evidence that the O-glycoproteome is differentially regulated and dynamic. The greatly expanded view of the O-glycoproteome should facilitate the exploration of how site-specific O-glycosylation regulates protein function.
U2 - 10.1038/emboj.2013.79
DO - 10.1038/emboj.2013.79
M3 - Journal article
C2 - 23584533
VL - 32
SP - 1478
EP - 1488
JO - E M B O Journal
JF - E M B O Journal
SN - 0261-4189
IS - 10
ER -