Immunochemical detection of protein dityrosine in atherosclerotic lesion of apo-E-deficient mice using a novel monoclonal antibody.
Dityrosine is one of the specific biomarkers for proteinoxidation. We prepared an antibody specific for proteindityrosine using a dimer of 3-p-(hydroxyphenyl)propionic acid (di-HP) as a hapten. Three clones (A8, G6, and 1C3) were obtained, and the antibody from the A8 clone reacted with the di-HP-conjugated protein but not with a dityrosine conjugate. The others (G6 and 1C3 clones) recognized both the di-HP and dityrosine conjugates. The antibodies reacted with peptidyl dityrosine, derived from Thr-Tyr-Ser, rather than the free dityrosine. The reactivity of the latter two antibodies with lens proteins oxidized by incubation with H(2)O(2)/Cu was in good accordance with the formation of the dityrosine-like fluorescence. Using the obtained monoclonal antibody, the immunopositive staining of atherosclerotic lesions in Apo E-deficient mice was confirmed by an immunohistochemical technique.
Kato
Y
Y
School of Humanities for Environmental Policy and Technology, Himeji Institute of Technology, Himeji, 670-0092, Japan.
Wu
X
X
Naito
M
M
Nomura
H
H
Kitamoto
N
N
Osawa
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001013:Aorta, Thoracic / Q000378:metabolism; D001057:Apolipoproteins E / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D001161:Arteriosclerosis / Q000378:metabolism* / Q000473:pathology; D001667:Binding, Competitive; D015415:Biomarkers / Q000032:analysis; D003459:Crystallins / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D016877:Oxidants / Q000378:metabolism; D010666:Phenylpropionates / Q000276:immunology / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000941:Antigens; D001057:Apolipoproteins E; D015415:Biomarkers; D003459:Crystallins; D006241:Haptens; D007106:Immune Sera; D016877:Oxidants; D010666:Phenylpropionates; D014443:Tyrosine; D006861:Hydrogen Peroxide; C007543:dityrosine