A sensitive gel-based global O-glycomics approach reveals high levels of mannosyl glycans in the high mass region of the mouse brain proteome
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Isabelle Breloy
,
Sandra Pacharra
Abstract
We developed a gel-based global O-glycomics method applicable for highly complex protein mixtures entrapped in discontinuous gradient gel layers. The protocol is based on in-gel proteolysis with pronase followed by (glyco)peptide elution and off-gel reductive β-elimination. The protocol offers robust performance with sensitivity in the low picomolar range, is compatible with gel-based proteomics, and shows superior performance in global applications in comparison with workflows eliminating glycans in-gel or from electroblotted glycoproteins. By applying this method, we analyzed the O-glycome of human myoblasts and of the mouse brain O-glycoproteome. After semipreparative separation of mouse brain proteins by one-dimensional SDS gel electrophoresis, the O-glycans from proteins in different mass ranges were characterized with a focus on O-mannose-based glycans. The relative proportion of the latter, which generally represent a rare modification, increases to comparatively high levels in the mouse brain proteome in dependence of increasing protein masses.
©2012 by Walter de Gruyter Berlin Boston
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Articles in the same Issue
- Masthead
- Masthead
- Guest Editorial
- Highlight: Dolce vita – a sample from European glycobiology
- Highlight: Dolce Vita – A Sample From European Glycobiology
- Complicated N-linked glycans in simple organisms
- Methylation – an uncommon modification of glycans
- Protein O-glycosylation analysis
- A sensitive gel-based global O-glycomics approach reveals high levels of mannosyl glycans in the high mass region of the mouse brain proteome
- Glycan profiles of the 27 N-glycosylation sites of the HIV envelope protein CN54gp140
- The analysis of N-glycans of cell membrane proteins from human hematopoietic cell lines reveals distinctions in their pattern
- The structure of the extracellular teichoic acids from the allergy-protective bacterium Lactococcus lactis G121
- Aminoquinolines as fluorescent labels for hydrophilic interaction liquid chromatography of oligosaccharides
- Fungal α-arabinofuranosidases of glycosyl hydrolase families 51 and 54 show a dual arabinofuranosyl- and galactofuranosyl-hydrolyzing activity
- Glycoengineering the N-acyl side chain of sialic acid of human erythropoietin affects its resistance to sialidase
- Shiga toxin glycosphingolipid receptor expression and toxin susceptibility of human pancreatic ductal adenocarcinomas of differing origin and differentiation
- Review
- Structural determinants of the eosinophil cationic protein antimicrobial activity
- Research Articles/Short Communications
- Membranes, Lipids, Glycobiology
- Anticancer peptide NK-2 targets cell surface sulphated glycans rather than sialic acids
- Proteolysis
- Extracellular aspartic protease SAP2 of Candida albicans yeast cleaves human kininogens and releases proinflammatory peptides, Met-Lys-bradykinin and des-Arg9-Met-Lys-bradykinin
