In vitro and in vivo characterization of plant-specific glycation products in terms of their structure, origin and effects on mammalian cells

• Applicant: Dr. Andrej Frolov
• Subject Area: Food Chemistry
• Term: from 2012 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 224903464

Glycation, i.e. the reaction of free amino groups with reducing sugars, yields Amadori and Heyns products that can undergo degradation and cross-linking, producing toxic, pro-inflammatory, allergenic, and diabetogenic advanced glycation end-products (AGEs). This phenomenon was recently shown to accompany plant environmental stress and will be studied here in more detail. First, the RuBisCO large subunit and selected peptides representing its sequence will be glycated/glycoxidated using various sugars and reactive carbonyls as modification agents. The products will be studied by tandem mass spectrometry to characterize their characteristic fragmentation patterns using also synthetic model peptides if required. Based on this data, highly sensitive LC-MS/MS-techniques and appropriate enrichment procedures will be developed to characterize glycation and glycoxidation in plants (A. thaliana) during drought and/or light stress. These stress conditions are highly relevant for many agricultural regions and might even gain more attention due to prognosed climate changes. Proteome alterations accompanying drought and light stress will reveal characteristic stress-related glycation and glycoxidation patterns. Additionally, RuBisCO will be isolated from stressed and nonstressed plants and its in vivo modification patterns will be compared with those obtained in vitro. Furthermore, the physiological effects of in vivo glycated/glycoxidated RuBisCO will be characterized.

• DFG Programme: Research Grants