Abstract
Key message
The identification of N -glycosylated proteins with information about changes in the level of N -glycosylation during de-etiolation provides a database that will aid further research on plant N -glycosylation and de-etiolation.
Abstract
N-glycosylation is one of the most prominent and abundant protein post-translational modifications in all eukaryotes and in plants it plays important roles in development, stress tolerance and immune responses. Because light-induced de-etiolation is one of the most dramatic developmental processes known in plants, seedlings undergoing de-etiolation are an excellent model for investigating dynamic proteomic profiles. Here, we present a comprehensive, quantitative N-glycoproteomic profile of maize seedlings undergoing 12 h of de-etiolation obtained using Concanavalin A (Con A) lectin affinity chromatography enrichment coupled with a nano-LC–MS/MS-based iTRAQ approach. In total, 1084 unique N-glycopeptides carrying 909 N-glycosylation sites and corresponding to 609 proteins were identified and quantified, including 186 N-glycosylation sites from 162 proteins that were significantly regulated over the course of the 12 h de-etiolation period. Based on hierarchical clustering analysis, the significantly regulated N-glycopeptides were divided into seven clusters that showed different N-glycosylation patterns during de-etiolation. We found no obvious difference in the enriched MapMan bincode categories for each cluster, and these clustered significantly regulated N-glycoproteins (SRNPs) are enriched in miscellaneous, protein, cell wall and signaling, indicating that although the N-glycosylation regulation patterns of these SRNPs might differ, they are involved in similar biological processes. Overall, this study represents the first large-scale quantitative N-glycoproteome of the model C4 plant, maize, which is one of the most important cereal and biofuel crops. Our results greatly expand the maize N-glycoproteomic database and also shed light on the potential roles of N-glycosylation modification during the greening of maize leaves.
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Abbreviations
- Con A:
-
Concanavalin A
- ER:
-
Endoplasmic reticulum
- GO:
-
Gene ontology
- iTRAQ:
-
Isobaric tags for relative and absolute quantification
- LAC:
-
Lectin affinity chromatography
- LC–MS/MS:
-
Liquid chromatography and tandem mass spectrometry
- RLK:
-
Receptor-like kinase
- SRNP:
-
Significantly regulated N-glycoprotein
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This study was funded by the “Strategic Priority Research Program” of the Chinese Academy of Sciences (Grant No. XDA08010206.5).
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Communicated by Mark C. Jordan.
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Fig. S1. Schematic of protein N-glycosylation in the secretory pathway. Enzymes identified as being N-glycosylated in this study are highlighted in yellow. (PDF 3978 kb)
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Bu, Tt., Shen, J., Chao, Q. et al. Dynamic N-glycoproteome analysis of maize seedling leaves during de-etiolation using Concanavalin A lectin affinity chromatography and a nano-LC–MS/MS-based iTRAQ approach. Plant Cell Rep 36, 1943–1958 (2017). https://doi.org/10.1007/s00299-017-2209-x
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DOI: https://doi.org/10.1007/s00299-017-2209-x