Qualitative determination of specific protein glycation products by matrix-assisted laser desorption/ionization mass spectrometry Peptide mapping.
The nonenzymatic reaction between reducing sugars and proteins, known as the Maillard reaction, has received increased recognition from nutritional science and medical research. The development of new analytical techniques for the detection of protein-bound Maillard products is therefore crucial. In this study, we applied peptide mapping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to investigate the formation of structurally specific Maillard products on glycated lysozyme (AGE-lysozyme), produced upon incubation with D-glucose. In parallel, we synthesized N(epsilon)-(carboxymethyl)lysine-modified lysozyme (CML-lysozyme) and N(epsilon)-(carboxyethyl)lysine-modified lysozyme, two well-described glycation products, as model substances. 3-Deoxyglucosone-modified lysozyme and methylglyoxal-modified lysozyme were prepared as examples of glycation products incubated with dicarbonyl compounds. We were able to detect specific modifications on AGE-lysozyme, which were assigned to CML, imidazolone A, and the Amadori product.
Humeny
Andreas
A
Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander Universität Erlangen-Nürnberg, Fahrstrasse 17, 91054 Erlangen, Germany.
Kislinger
Thomas
T
Becker
Cord-Michael
CM
Pischetsrieder
Monika
M
D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005947:Glucose / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis*; D006031:Glycosylation; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D015416:Maillard Reaction; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D010449:Peptide Mapping; D011765:Pyruvaldehyde / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013329:Structure-Activity Relationship
D006023:Glycoproteins; C048496:N(6)-carboxymethyllysine; D011765:Pyruvaldehyde; D003847:Deoxyglucose; D009113:Muramidase; C016350:3-deoxyglucosone; D005947:Glucose; D008239:Lysine