Lectin affinity-based glycoproteome analysis of the developing xylem in poplar

Abstract

Glycosylation is a significant post-translational modification of the proteins, and some glycoproteins serve as the players in plant cell wall synthesis and modification. Wood is a highly developed cell wall organization, and protein glycosylation as a regulatory way might be involved in wood formation. Here, a lectin affinity-based glycoproteome was performed in stem developing xylem of poplar. After enrichment, trypsin digestion, LC-MS/MS analysis and peptide identification, we identified 154 glycoproteins from poplar developing xylem, which were classified into 9 functional groups mainly including protein acting on carbohydrates, oxido-reductase, proteases, and protein kinases. Further, N- and/or O-glycosylation sites of the identified proteins were analyzed by the bioinformatic tools, and deglycosylation experiments in the selected PtSOD and PtHAD proteins verified the reliability of the identified glycoproteins. Analysis of protein subcellular localization showed that total 63 percent of the identified glycoproteins were extracellular proteins or located in the plasma membrane. Poplar eFP and RT-qPCR data showed that a number of the genes encoding these glycoproteins such as laccase, peroxidase and cysteine protease, have highly preferential expression profiles in the developing xylem. Together with the literatures, most identified glycoproteins could be involved in wood cell wall synthesis and modification in poplar. Thus, our study provides some potential wood formation-related glycoproteins to be determined during tree stem development.