Maillard reaction-like lysine modification by a lipid peroxidation product: immunochemical detection of protein-bound 2-hydroxyheptanal in vivo.
2-Hydroxyheptanal (2-HH) is one of the reactive aldehyde species generated during the peroxidation of n-6 polyunsaturated fatty acids, such as linoleic and arachidonic acids. Analogous to the Maillard reaction of reducing sugars, 2-HH readily reacts with lysine epsilon-amino groups. In the present study, to define the occurrence of the Maillard reaction-like lysine modification by 2-HH in vivo, we raised a monoclonal antibody directed to a trihydropyridinone (THPO) structure, 1-alkyl-4-butyl-5-pentyl-1,2,6-trihydropyridin-3-one, formed from 2-HH and lysine, and examined the presence of the antigenic structure in the human atherosclerotic aorta. Mice were immunized with the 2-HH-modified keyhole limpet hemocyanin (KLH) as the immunogen. Using a THPO-carrier protein conjugate, we screened the hybridomas and finally obtained a clone that produced the monoclonal antibody 3C8 (mAb3C8). The antibody strongly recognized bovine serum albumin (BSA) treated with 2-HH, but showed no cross-reactivity with BSAs modified with other related aldehydes. By using this antibody, it was revealed that the antigenic structure was indeed present in atherosclerotic lesions of the human aorta.
Itakura
Koichi
K
Faculty of Education, Aichi University of Education, Kariya, Japan. kitakura@auecc.aichi-edu.ac.jp
Furuhata
Atsunori
A
Shibata
Noriyuki
N
Kobayashi
Makio
M
Uchida
Koji
K
D000447:Aldehydes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001011:Aorta / Q000737:chemistry; D001161:Arteriosclerosis / Q000378:metabolism* / Q000473:pathology; D002208:Caproates / Q000737:chemistry / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D005260:Female; D006241:Haptens; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D007120:Immunochemistry; D015227:Lipid Peroxidation; D008239:Lysine / Q000737:chemistry; D015416:Maillard Reaction; D051379:Mice; D008807:Mice, Inbred BALB C; D008956:Models, Chemical; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D011728:Pyridones / Q000737:chemistry / Q000276:immunology
C477439:1-carboxypentyl-4-butyl-5-pentyl-1,2,6-trihydropyridin-3-one; D000447:Aldehydes; D000911:Antibodies, Monoclonal; D002208:Caproates; D000939:Epitopes; D006241:Haptens; D011506:Proteins; D011728:Pyridones; C098838:2-hydroxyheptanal; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D008239:Lysine