CHARACTERIZATION OF A SPECIFIC POLYCLONAL ANTIBODY AGAINST 13-HYDROPEROXYOCTADECADIENOIC ACID-MODIFIED PROTEIN - FORMATION OF LIPID HYDROPEROXIDE-MODIFIED APO-B-100 IN OXIDIZED LDL

Citation
Y. Kato et al., CHARACTERIZATION OF A SPECIFIC POLYCLONAL ANTIBODY AGAINST 13-HYDROPEROXYOCTADECADIENOIC ACID-MODIFIED PROTEIN - FORMATION OF LIPID HYDROPEROXIDE-MODIFIED APO-B-100 IN OXIDIZED LDL, Journal of lipid research, 38(7), 1997, pp. 1334-1346
Citations number
51
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology
Journal title
ISSN journal
0022-2275
Volume
38
Issue
7
Year of publication
1997
Pages
1334 - 1346
Database
ISI
SICI code
0022-2275(1997)38:7<1334:COASPA>2.0.ZU;2-C
Abstract
Lipid hydroperoxide may react with protein or amino phospholipid witho ut secondary decomposition. We prepared a polyclonal antibody to lipid hydroperoxide-modified proteins using 13S-hydroperoxy-9Z, 11E-octadec adienoic acid-modified keyhole limpet hemocyanin (13-HPODE-KLH) as imm unogen. The antibody recognized 13-HPODE-modified bovine serum albumin (BSA), but not aldehyde-modified proteins, such as malondialdehyde-mo dified BSA. The antibody also recognized adducts derived from 13-HPODE and 13S- hydroperoxy-9Z, 11E, 15Z-octadecatrienoic acid (13-HPOTRE(al pha)). The oxidized alpha-linolenic acid- and linoleate-protein adduct s were recognized by the antibody. Oxidized phospholipid-protein adduc ts were scarcely recognized by the antibody. However, when ester bonds of phospholipids containing linoleic acid were hydrolyzed by alkaline treatment, the cross-reactivities appeared. The result suggests that a phospholipid hydroperoxide can react with a protein directly or indi rectly, and a carboxyl terminal (COOH) of the lipid in an adduct was n eeded as an epitope. Oxidized LDL (ox-LDL) was prepared by the incubat ion of LDL with copper ion or 2,2'-azobis (2-amidinopropane) dihydroch loride (AAPH), and the formation of lipid hydroperoxide-modified apoli poprotein was confirmed using the antibody. A slight immunoreactivity was observed in ox-LDL without alkaline treatment. When the ox-LDL was treated with alkali to hydrolyze the ester bonds of the lipid, enhanc ed antigenicity appeared with time-dependency. The results suggest tha t lipid hydroperoxide-modified apolipoprotein was formed during the ox idation of LDL.