In-depth glycoproteomic characterisation of grape berry vacuolar invertase using a combination of mass spectrometry-based approaches.
Fiche publication
Date publication
juin 2016
Journal
Food chemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Mme SCHAEFFER-REISS Christine, Dr VAN DORSSELAER Alain
Tous les auteurs :
Hovasse A, Alayi TD, Van Dorsselaer A, Marchal R, Jégou S, Schaeffer-Reiss C
Lien Pubmed
Résumé
Vacuolar invertase is a key enzyme of sugar metabolism in grape berries. A full characterisation of this highly N-glycosylated protein is required to help understand its biological and biochemical significance in grapes. We have developed a mass spectrometry (MS)-based glycoproteomic approach wherein deglycosylated peptides are analysed by LC-MS/MS, while intact glycopeptides are characterised using a dedicated MS method to determine the attachment sites and micro-heterogeneity. For grape invertase, in parallel with deglycosylated peptides analysis, different enzymatic digestions were performed and glycopeptide detection was improved by enrichment method, nanoLC-MS and oxonium glycan ions. This MS-based glycoproteomic approach demonstrates that vacuolar invertase is glycosylated at all twelve potential N-glycosylation sites. Glycosylation is heterogeneous, with twelve glycoforms identified at six of the sites. The identification of several types of N-glycans is a major result to correlate with the surface and foaming properties of wine, the solubility, allergenicity, and protease resistance of wine proteins.
Mots clés
Amino Acid Sequence, Glycopeptides, analysis, Molecular Sequence Data, Tandem Mass Spectrometry, methods, Vitis, enzymology, beta-Fructofuranosidase, chemistry
Référence
Food Chem. 2016 Jun;200:237-44