Immunochemical Detection of Carboxymethylated Apo B-100 in Copper-Oxidized LDL

Kato, Yoji; Tokunaga, Katsuhiko; Osawa, Toshihiko

doi:10.1006/bbrc.1996.1451

Cited by 18 publications

(7 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…N α ‐ p ‐Tosyl‐ N ϵ ‐(carboxymethyl)‐ l ‐lysine‐methyl ester (Tos‐CML‐Me) was synthesized as reported by us [21]. CML derivative of BSA (CML‐BSA) was synthesized as reported by Kato et al [22]. Pentosidine was synthesized according to the method of Sell and Monnier [23].…”

Section: Methodsmentioning

confidence: 99%

Imidazolone, a novel advanced glycation end product, is present at high levels in kidneys of rats with streptozotocin‐induced diabetes

et al. 1997

View full text Add to dashboard Cite

We produced a monoclonal antibody to imidazolones A and B, novel advanced glycation end products formed from the reaction of 3-deoxyglucosone (3-DG) with the guanidino group of arginine. Liquid chromatography/mass spectrometry demonstrated that the formation of imidazolone A by incubating 3-DG with arginine is very rapid, reaching a maximum concentration within 24 h, but the formation of imidazolone B is very slow and low in quantity even after 2 weeks. Thus, at physiological conditions the formation of imidazolone A is dominant, while that of imidazolone B is negligible. Immunochemistry demonstrated that the imidazolone content in the kidneys of streptozotocin-induced diabetic rats was significantly higher than in the control rats. Serum levels of 3-DG in the diabetic rats were also significantly higher than in control rats. 3-DG attacks the arginine residues of the tissue proteins, producing imidazolone at high levels in the kidneys affected by diabetic nephropathy.

show abstract

Section: Methodsmentioning

confidence: 99%

Imidazolone, a novel advanced glycation end product, is present at high levels in kidneys of rats with streptozotocin‐induced diabetes

et al. 1997

View full text Add to dashboard Cite

show abstract

“…Modification of LDL-Human LDL was isolated from healthy volunteers using density centrifugation (15). The modification of LDL was performed by copper ion and 2,2Ј-azobis(2-amidinopropane)dihydrochloride (AAPH) as described previously (9).…”

Section: Synthesis Of N-acetyl-glycyl-nmentioning

confidence: 99%

Formation of N ε-(Hexanonyl)lysine in Protein Exposed to Lipid Hydroperoxide

Kato

Mori

Makino

et al. 1999

Journal of Biological Chemistry

Self Cite

161

View full text Add to dashboard Cite

The objectives of this study were to estimate the structure of the lipid hydroperoxide-modified lysine residue and to prove the presence of the adducts in vivo. The reaction of lipid hydroperoxide toward the lysine moiety was investigated employing N-benzoyl-glycyl-L-lysine (Bz-Gly-Lys) as a model compound of Lys residues in protein and 13-hydroperoxyoctadecadienoic acid (13-HPODE) as a model of the lipid hydroperoxides. One of the products, compound X, was isolated from the reaction mixture of 13-HPODE and Bz-Gly-Lys and was then identified as N-benzoyl-glycyl-N ⑀ -(hexanonyl)lysine. To prove the formation of N ⑀ -(hexanonyl)lysine, named HEL, in protein exposed to the lipid hydroperoxide, the antibody to the synthetic hexanonyl protein was prepared and then characterized in detail. Using the anti-HEL antibody, the presence of HEL in the lipid hydroperoxide-modified proteins and oxidized LDL was confirmed. Furthermore, the positive staining by anti-HEL antibody was observed in human atherosclerotic lesions using an immunohistochemical technique. The amide-type adduct may be a useful marker for the lipid hydroperoxide-derived modification of biomolecules.During lipid peroxidation, biomolecules such as proteins or aminolipids can be covalently modified by lipid decomposition products. For the case of aliphatic aldehydes (alkanals) such as 1-hexanal or 1-nonanal, the N ⑀ -amino groups of the lysine residues in protein can be modified through the formation of a Schiff base. ␣,␤-Unsaturated aldehydes (alkenals) such as acrolein or 4-hydroxy-2-nonenal react with lysine, cysteine, and histidine through a Michael-type addition (1, 2). On the other hand, lipid hydroperoxide might covalently react with protein without serious decomposition of its structure (3). Keto fatty acid (4), which is one of the products by lipoxygenase reaction, can also react with protein and amino acids as previously suggested (5-7). In addition, the pyrrole compounds from long chain epoxides and lysine were identified (8). However, the mechanism of lipid hydroperoxide-derived protein modification is not so clear.To estimate the structure after lipid hydroperoxide-derived lysine modification, the reaction of 13-hydroperoxyoctadecadienoic acid 1 with N-benzoyl-glycyl-L-lysine (BzGly-Lys) was investigated. In this study, a novel compound, N-benzoyl-glycyl-N ⑀ -(hexanonyl)lysine (named HEL), was identified as one of the lipid hydroperoxide-modified lysine residues. The formation of HEL in lipid hydroperoxide-modified proteins including oxidatively modified LDL was confirmed using the specific antibody to the HEL residue. In addition, the HEL moiety was detected in human atherosclerotic plaques by immunohistochemical approach. As far as we know, the formation of an amide-type adduct has not been previously reported. This novel adduct derived from lipid hydroperoxide may become an initial marker for the oxidative damage of biological molecules in vivo. EXPERIMENTAL PROCEDURESMaterials-The chemicals used were from the following sources. Bz-Gly-Lys an...

show abstract

“…Aldehyde-modified BSAs were prepared using malondialdehyde, 1-hexanal, and 2-hexenal, and synthetic 4-hydroxy-2-nonenal as described previously (19). Peroxidized lipidmodified BSA, lipid hydroperoxide-modified BSA, and the modified BSA having various alkyl chain lengths in the amide-type adduct were prepared as described (6).…”

Section: Preparation Of Chemically-modified Proteins and Nmentioning

confidence: 99%