Details

Autor(en) / Beteiligte

• Steentoft, Catharina
• Vakhrushev, Sergey Y
• Joshi, Hiren J
• Kong, Yun
• Vester-Christensen, Malene B
• Schjoldager, Katrine T-B G
• Lavrsen, Kirstine
• Dabelsteen, Sally
• Pedersen, Nis B
• Marcos-Silva, Lara
• Gupta, Ramneek
• Paul Bennett, Eric
• Mandel, Ulla
• Brunak, Søren
• Wandall, Hans H
• Levery, Steven B
• Clausen, Henrik

Titel

Precision mapping of the human O-GalNAc glycoproteome through SimpleCell technology

Ist Teil von

• The EMBO journal, 2013-05, Vol.32 (10), p.1478-1488

Ort / Verlag

Chichester, UK: John Wiley & Sons, Ltd

Erscheinungsjahr

2013

Link zum Volltext

Quelle

Wiley Online Library - AutoHoldings Journals

Beschreibungen/Notizen

• 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. Comprehensive proteomics survey in 12 human cell lines and development of an improved NetOGlyc4.0 prediction tool greatly expand the view on mucin‐type protein O‐glycosylation.