title: A Snapshot of the Plant Glycated Proteome: STRUCTURAL, FUNCTIONAL, AND MECHANISTIC ASPECTS.
abstract: Glycation is the reaction of carbonyl compounds (reducing sugars and α-dicarbonyls) with amino acids, lipids, and proteins, yielding early and advanced glycation end products (AGEs). The AGEs can be formed via degradation of early glycation intermediates (glycoxidation) and by interaction with the products of monosaccharide autoxidation (autoxidative glycosylation). Although formation of these potentially deleterious compounds is well characterized in animal systems and thermally treated foods, only a little information about advanced glycation in plants is available. Thus, the knowledge of the plant AGE patterns and the underlying pathways of their formation are completely missing. To fill this gap, we describe the AGE-modified proteome ofBrassica napusand characterize individual sites of advanced glycation by the methods of liquid chromatography-based bottom-up proteomics. The modification patterns were complex but reproducible: 789 AGE-modified peptides in 772 proteins were detected in two independent experiments. In contrast, only 168 polypeptides contained early glycated lysines, which did not resemble the sites of advanced glycation. Similar observations were made withArabidopsis thaliana The absence of the early glycated precursors of the AGE-modified protein residues indicated autoxidative glycosylation, but not glycoxidation, as the major pathway of AGE formation. To prove this assumption and to identify the potential modifying agents, we estimated the reactivity and glycative potential of plant-derived sugars using a model peptide approach and liquid chromatography-mass spectrometry-based techniques. Evaluation of these data sets together with the assessed tissue carbohydrate contents revealed dihydroxyacetone phosphate, glyceraldehyde 3-phosphate, ribulose, erythrose, and sucrose as potential precursors of plant AGEs.
authors.affiliation: From the Departments of Bioorganic Chemistry and Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Bilova
authors.affiliation: Departments of Biochemistry and Plant Physiology and Biochemistry, Faculty of Biology, Saint Petersburg State University, 199034 Saint Petersburg, Russia.
authors.forename: Elena
authors.initials: E
authors.lastname: Lukasheva
authors.affiliation: Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), D-06466 Stadt Seeland, Germany, and.
authors.forename: Dominic
authors.initials: D
authors.lastname: Brauch
authors.affiliation: Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany.
authors.forename: Uta
authors.initials: U
authors.lastname: Greifenhagen
authors.affiliation: From the Departments of Bioorganic Chemistry and Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany.
authors.forename: Gagan
authors.initials: G
authors.lastname: Paudel
authors.affiliation: Plant Physiology and Biochemistry, Faculty of Biology, Saint Petersburg State University, 199034 Saint Petersburg, Russia.
authors.forename: Elena
authors.initials: E
authors.lastname: Tarakhovskaya
authors.affiliation: Interdisciplinary Center for Crop Plant Research (IZN), Martin Luther University Halle-Wittenberg, D-06120 Halle (Saale),Germany.
authors.forename: Nadezhda
authors.initials: N
authors.lastname: Frolova
authors.affiliation: Interdisciplinary Center for Crop Plant Research (IZN), Martin Luther University Halle-Wittenberg, D-06120 Halle (Saale),Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Mittasch
authors.affiliation: Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry (IPB), D-06120 Halle (Saale), Germany.
authors.forename: Gerd Ulrich
authors.initials: GU
authors.lastname: Balcke
authors.affiliation: Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry (IPB), D-06120 Halle (Saale), Germany.
authors.forename: Alain
authors.initials: A
authors.lastname: Tissier
authors.affiliation: Plant Physiology and Biochemistry, Faculty of Biology, Saint Petersburg State University, 199034 Saint Petersburg, Russia.
authors.forename: Natalia
authors.initials: N
authors.lastname: Osmolovskaya
authors.affiliation: Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry (IPB), D-06120 Halle (Saale), Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vogt
authors.affiliation: From the Departments of Bioorganic Chemistry and.
authors.forename: Ludger A
authors.initials: LA
authors.lastname: Wessjohann
authors.affiliation: Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Birkemeyer
authors.affiliation: Interdisciplinary Center for Crop Plant Research (IZN), Martin Luther University Halle-Wittenberg, D-06120 Halle (Saale),Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Milkowski
authors.affiliation: From the Departments of Bioorganic Chemistry and Faculty of Chemistry and Mineralogy, Universität Leipzig, D-04103 Leipzig, Germany, afrolov@ipb-halle.de.
authors.forename: Andrej
authors.initials: A
authors.lastname: Frolov
mesh_terms: D029688:Brassica napus / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics
chemical_list: D006023:Glycoproteins; D010940:Plant Proteins; D020543:Proteome
keywords: Arabidopsis thaliana; Brassica napus; advanced glycation end products (AGEs); carbohydrate-binding protein; gas chromatography-mass spectrometry (GC-MS); glycation; mass spectrometry (MS); metabolomics; plant biochemistry; proteomics