Immunological evidence for methylglyoxal-derived modifications in vivo. Determination of antigenic epitopes.
The Maillard reaction, a non-enzymatic reaction of ketones and aldehydes with amino groups of proteins, contributes to the aging of proteins and to complications associated with diabetes. Methylglyoxal (MG) is a 2-oxoaldehyde derived from glycolytic intermediates and produced during the Maillard reaction. We reported previously the formation of a lysine-lysine protein cross-linking structure (imidazolysine) and a fluorescent arginine modification (argpyrimidine) from the Maillard reaction of MG. Here we show that rabbit antibodies to MG-modified ribonuclease A identify proteins modified by the Maillard reaction of glucose, fructose, ribose, glyceraldehyde, glyoxal, ascorbate, and ascorbate oxidation products (dehydroascorbate, 2,3-diketogulonate, L-xylosone, and L-threose) in addition to those modified by MG. The antibody recognized imidazolysine and argpyrimidine and a glyoxal-derived lysine-lysine cross-link. It did not react with Nepsilon-carboxymethyllysine. Incubations with amino acids revealed strongest reactivity with Nalpha-t-butoxycarbonylarginine and MG, and we identified argpyrimidine as one of the epitopes from this incubation mixture. Serum proteins from human diabetics reacted more strongly with the antibody than those from normal individuals, and the levels correlated with glycemic control. Collagen from human corneas contained MG-derived modifications, with those from older subjects containing higher levels of modified proteins than those from younger ones. An immunoaffinity-purified antibody showed higher reactivity with old corneas than with younger ones and localized the antigens primarily within the stromal region of the cornea. These results confirm reported MG-derived modifications in tissue proteins and show that dicarbonyl-mediated protein modification occurs during Maillard reactions in vivo.
Shamsi
F A
FA
Center for Vision Research, Department of Ophthalmology, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Partal
A
A
Sady
C
C
Glomb
M A
MA
Nagaraj
R H
RH
D000328:Adult; D000368:Aged; D000375:Aging / Q000276:immunology; D000818:Animals; D001798:Blood Proteins / Q000276:immunology*; D003920:Diabetes Mellitus / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D017127:Glycation End Products, Advanced; D006801:Humans; D015416:Maillard Reaction; D008875:Middle Aged; D011765:Pyruvaldehyde / Q000494:pharmacology*; D011817:Rabbits
D001798:Blood Proteins; D000939:Epitopes; D017127:Glycation End Products, Advanced; D011765:Pyruvaldehyde