Combining various strategies to increase the coverage of the plant cell wall glycoproteome.
Glycoproteomics recently became a very active field, mostly in mammals. The first part of this paper consists of a mini-review on the strategies used in glycoproteomics, namely methods for enrichment in glycoproteins and mass spectrometry (MS) techniques currently used. In a second part, these strategies are applied to the cell wall glycoproteome of etiolated hypocotyls of Arabidopsis thaliana, showing their complementarity. Several sub-glycoproteomes were obtained by: (i) affinity chromatography on concanavaline A (ConA) and analysis of glycoproteins by MALDI-TOF MS; (ii) multidimensional lectin chromatography (using AIL, PNA, ConA and WGA lectins) and subsequent identification of glycoproteins by MALDI-TOF MS and LC-MS/MS; (iii) boronic acid chromatography followed by identification of glycoproteins by MALDI-TOF MS. Altogether, 127 glycoproteins were identified. Most glycoproteins were found to be putative N-glycoproteins and N-glycopeptides were predicted from MS data using the ProTerNyc bioinformatics software.
Université de Toulouse, UPS, UMR 5546, Surfaces Cellulaires et Signalisation chez les Végétaux, BP 42617, F-31326 Castanet-Tolosan, France.
Yu
Y
Zhang
Aurélie
A
Giboulot
Michel
M
Zivy
Benoît
B
Valot
Elisabeth
E
Jamet
Cécile
C
Albenne
D017360:Arabidopsis / Q000737:chemistry* / Q000166:cytology*; D029681:Arabidopsis Proteins / Q000032:analysis*; D002473:Cell Wall / Q000737:chemistry*; D006023:Glycoproteins / Q000032:analysis*; D040901:Proteomics
D029681:Arabidopsis Proteins; D006023:Glycoproteins