title: Maillard reaction-like lysine modification by a lipid peroxidation product: immunochemical detection of protein-bound 2-hydroxyheptanal in vivo.
abstract: 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.
authors.affiliation: Faculty of Education, Aichi University of Education, Kariya, Japan. kitakura@auecc.aichi-edu.ac.jp
authors.forename: Koichi
authors.initials: K
authors.lastname: Itakura
authors.forename: Atsunori
authors.initials: A
authors.lastname: Furuhata
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Shibata
authors.forename: Makio
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Koji
authors.initials: K
authors.lastname: Uchida
mesh_terms: 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
chemical_list: 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