Protein modification by a Maillard reaction intermediate methylglyoxal

FEBS Letters

Miyata, Toshio¹  Uchida, Koji²  Osawa, Toshihiko²  Oya, Tomoko²  Yasuda, Yoshinari¹  Khor, Ooi Thiang² 

[1] Institute of Medical Sciences and Department of Medicine, Tokai University School of Medicine, Isehara 259-11, Japan;Laboratory of Food and Biodynamics, Nagoya University School of Agricultural Sciences, Nagoya 464-01, Japan

关键词: Methylglyoxal;    Maillard reaction;    5-Methylimidazolone;    Antibody;    MG;    methylglyoxal;    AGEs;    advanced glycation end products;    CML;    Nϵ -(carboxymethyl)lysine;    KLH;    keyhole limpet hemocyanin;    ELISA;    enzyme-linked immunosorbent assay;    HEPES;    N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid;    BSA;    bovine serum albumin;    TBS;    Tris-buffered saline;   

DOI  :  10.1016/S0014-5793(97)00610-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Methylglyoxal (MG), an endogenous metabolite that increases in diabetes, is a common intermediate in nonenzymatic glycation (Maillard reaction) in vivo. Here we describe the immunochemical approach to the detection of MG adducts in proteins in vitro and in atherosclerotic lesions of human aorta in vivo. The reaction of protein (bovine serum albumin) with MG led to selective loss of arginine and lysine residues, accompanied by the formation of 5-methylimidazolone (N ^δ -(5-methylimidazolon-2-yl)ornithine) and imidazolysine (1,3-di-lysino-4-methylimidazole) derivatives, respectively. The anti-5-methylimidazolone antibody was prepared by immunizing rabbits with a MG-keyhole limpet hemocyanin conjugate and purifying the serum on an affinity gel prepared by covalent attachment of the 5-methylimidazolone derivative. The antibody cross-reacted with the proteins treated with not only MG but trioses, such as hydroxyacetone, dihydroxyacetone, and glyceraldehyde. The immunohistochemical analysis revealed that atherosclerotic lesions of human aorta contained 5-methylimidazolone derivatives whose distributions were identical to those of advanced glycation end products (AGEs) detected by the anti-AGE antibody.

【 授权许可】

Unknown