TY - GEN

T1 - Probing of protein glycation by peptide-based model systems: analysis of peptide products in parallel to sugar and α-dicarbonyl intermediates

AU - Фролова, Надежда Владимировна

AU - Herfurth, Uta M.

AU - Nguyen, Duc Viet

AU - Soboleva, Alena

AU - Balcke, Gerd Ulrich

AU - Birkemeyer, Claudia

AU - Frolov, Andrej

N1 - Conference code: 6

PY - 2019

Y1 - 2019

N2 - Glycation is a non-enzymatic modification formed by reaction of reducing sugars with amino groups of proteins. Resulting Amadori compounds oxidize ('glycoxidation'), yielding advanced glycation end-products (AGEs), a heterogeneous group of potentially pro-inflammatory compounds. Alternatively, carbohydrates are involved in formation of α-dicarbonyls, yielding AGEs upon reaction with lysyl and arginyl residues ('oxidative glycosylation'). Despite the well-known deteriorating effect of AGEs, the exact formation mechanisms, relative contribution and possible interference of these pathways are unknown. Therefore, here we address glycation potential of dietary sugars and estimate the contribution of the major glycation pathways in formation of corresponding AGEs. Our experiments relied on model glycation systems, based on synthetic peptides, their Amadori modified counterparts and selected dietary sugar (or, in some cases, their 13C-labeled analogs). Thereby, all AGE-formation pathways can be considered simultaneously. Analysis of peptide, sugar and α-dicarbonyl intermediates will rely on RP-UHPLC-ESI-QqTOF-, GC-Q-EI- and RP-HPLC-IT-MS, in two latter cases, after appropriate derivatization. The structures of peptide products and sugar intermediates were identified by their MS/MS fragmentation patterns, whereas annotation of carbonyl compounds relied on EI patterns and co-elution with authentic standards. Individual glycation pathways were assigned by isotopic composition of products. Analysis of the glycation mixtures revealed 37 lysine- and 22 argininederived products. Annotation of the glycation pathways, assigned to the generation of lysine-derived AGEs, revealed their formation via several routes: (i) both glycoxidative and autoxidative pathways (α-amino semiadipic aldehyde-containing product and peptide fragments), (ii) mostly via “glycoxidation”, i.e. Amadori degradation (carboxymethylated peptides) or (iii) “autoxidative glycosylation” (pyrraline). This data were supported by kinetics profile of approximately 30 carbonyl compounds. To summarize, our approach allowed identification of the major routes for formation of specific AGEs. This project was supported by Deutsche Forschungsgemeinschaft (grant number FR3117/2-3) and RFBR (research project number 18-34-00927).

AB - Glycation is a non-enzymatic modification formed by reaction of reducing sugars with amino groups of proteins. Resulting Amadori compounds oxidize ('glycoxidation'), yielding advanced glycation end-products (AGEs), a heterogeneous group of potentially pro-inflammatory compounds. Alternatively, carbohydrates are involved in formation of α-dicarbonyls, yielding AGEs upon reaction with lysyl and arginyl residues ('oxidative glycosylation'). Despite the well-known deteriorating effect of AGEs, the exact formation mechanisms, relative contribution and possible interference of these pathways are unknown. Therefore, here we address glycation potential of dietary sugars and estimate the contribution of the major glycation pathways in formation of corresponding AGEs. Our experiments relied on model glycation systems, based on synthetic peptides, their Amadori modified counterparts and selected dietary sugar (or, in some cases, their 13C-labeled analogs). Thereby, all AGE-formation pathways can be considered simultaneously. Analysis of peptide, sugar and α-dicarbonyl intermediates will rely on RP-UHPLC-ESI-QqTOF-, GC-Q-EI- and RP-HPLC-IT-MS, in two latter cases, after appropriate derivatization. The structures of peptide products and sugar intermediates were identified by their MS/MS fragmentation patterns, whereas annotation of carbonyl compounds relied on EI patterns and co-elution with authentic standards. Individual glycation pathways were assigned by isotopic composition of products. Analysis of the glycation mixtures revealed 37 lysine- and 22 argininederived products. Annotation of the glycation pathways, assigned to the generation of lysine-derived AGEs, revealed their formation via several routes: (i) both glycoxidative and autoxidative pathways (α-amino semiadipic aldehyde-containing product and peptide fragments), (ii) mostly via “glycoxidation”, i.e. Amadori degradation (carboxymethylated peptides) or (iii) “autoxidative glycosylation” (pyrraline). This data were supported by kinetics profile of approximately 30 carbonyl compounds. To summarize, our approach allowed identification of the major routes for formation of specific AGEs. This project was supported by Deutsche Forschungsgemeinschaft (grant number FR3117/2-3) and RFBR (research project number 18-34-00927).

KW - protein glycation

KW - model peptides

KW - mass-spectrometry

KW - advanced glycation end-products (AGEs)

KW - sugars

KW - α-dicarbonyls

UR - https://pure.spbu.ru/admin/files/48615932/_2_II_1_6_2019_.pdf

M3 - Conference contribution

SN - 9785001505198

SN - 9785001505211 (Т. 2)

VL - 2

T3 - ACTA NATURAE (РУССКОЯЗЫЧНАЯ ВЕРСИЯ)

SP - 276

BT - II Объединенный научный форум: VI Съезд биохимиков России и IX Российский симпозиум «Белки и пептиды»

PB - Издательство «Перо»

CY - М.

Y2 - 1 October 2019 through 6 October 2019

ER -