Protein modification by a Maillard reaction intermediate methylglyoxal: Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo

Koji Uchida, Ooi Thiang Khor, Tomoko Oya, Toshihiko Osawa, Yoshinari Yasuda, Toshio Miyata

Research output: Contribution to journalArticlepeer-review

92 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)313-318
Number of pages6
JournalFEBS Letters
Volume410
Issue number2-3
DOIs
Publication statusPublished - 1997 Jun 30

Keywords

  • 5-Methylimidazolone
  • Antibody
  • Maillard reaction
  • Methylglyoxal

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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