Publication Information
Ma et al., 2016
No external accession available
Abstract
New Phytol. 2016 Oct;212(1):282-96. doi: 10.1111/nph.14014. Epub 2016 May 12.
Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated
glycoproteins in response to chilling stress in Arabidopsis.
Ma J(1), Wang D(1), She J(1)(2), Li J(1), Zhu JK(1), She YM(1).
Author information:
(1)Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences,
Shanghai, 201602, China.
(2)Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8,
Canada.
N-glycosylation has a great impact on glycoprotein structure, conformation,
stability, solubility, immunogenicity and enzyme activity. Structural
characterization of N-glycoproteome has been challenging but can provide
insights into the extent of protein folding and surface topology. We describe a
highly sensitive proteomics method for large-scale identification and
quantification of glycoproteins in Arabidopsis through (15) N-metabolic
labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis
and automated database searching. In-house databases of Arabidopsis
glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were
constructed by reducing 20% and 90% of the public database size, respectively,
to enable a rapid analysis of large datasets for comprehensive identification
and quantification of glycoproteins and heterogeneous N-glycans in a complex
mixture. Proteome-wide analysis identified c. 100 stress-related
N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were
examined to be up-regulated. Quantitative measurements provided a molecular
signature specific to glycoproteins for determining the degree of plant stress
at low temperature. Structural N-glycoproteomics following time-course cold
treatments revealed the stress-responsive degradation of high-mannose type
N-glycans in ER in response to chilling stress, which may aid in elucidating the
cellular mechanisms of protein relocation, transport, trafficking, misfolding
and degradation under stress conditions.
© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.
DOI: 10.1111/nph.14014
PMCID: PMC5513495
PMID: 27558752 [Indexed for MEDLINE]
