title: Amyloid beta 2-microglobulin is modified with N epsilon-(carboxymethyl)lysine in dialysis-related amyloidosis.
abstract: Recent work from this laboratory revealed that advanced glycation end product was localized to amyloid deposits in patients with dialysis-related amyloidosis by immunohistochemistry using a monoclonal antibody to advanced glycation end product. To elucidate the epitope of the antibody, N alpha-p-tosyl-L-lysine-methyl ester was incubated with glucose in vitro, and then a compound reactive to the antibody was purified from the incubate by buthanol extraction, XAD-2 column chromatography, and high-performance liquid chromatography while the reactivity was examined by enzyme linked immunosorbent assay. The purified compound was identified as N epsilon-(carboxymethyl)-N alpha-p-tosyl-L-lysine-methyl ester by using secondary ion mass spectrometry, and 1H- and 13C-nuclear magnetic resonance spectroscopy. The epitope of the antibody was identified as -CH2-NH-CH2-COOH by enzyme-linked immunosorbent assay of compounds with structures similar to N epsilon-(carboxymethyl)lysine. Immunochemical study using the antibody demonstrated the presence of N epsilon-(carboxymethyl)lysine in the beta 2-microglobulin dimer (molecular weight 23929) isolated from the synovium amyloid of a hemodialysis patient with dialysis-related amyloidosis. In conclusion, amyloid beta 2-microglobulin is modified with N epsilon-(carboxymethyl)lysine in dialysis-related amyloidosis.
authors.affiliation: Nagoya University Branch Hospital, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Niwa
authors.forename: M
authors.initials: M
authors.lastname: Sato
authors.forename: T
authors.initials: T
authors.lastname: Katsuzaki
authors.forename: T
authors.initials: T
authors.lastname: Tomoo
authors.forename: T
authors.initials: T
authors.lastname: Miyazaki
authors.forename: N
authors.initials: N
authors.lastname: Tatemichi
authors.forename: Y
authors.initials: Y
authors.lastname: Takei
authors.forename: T
authors.initials: T
authors.lastname: Kondo
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000686:Amyloidosis / Q000209:etiology*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003956:Dialysis / Q000009:adverse effects; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D017127:Glycation End Products, Advanced / Q000276:immunology*; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D001613:beta 2-Microglobulin / Q000737:chemistry*
chemical_list: D000682:Amyloid; D000939:Epitopes; D017127:Glycation End Products, Advanced; D001613:beta 2-Microglobulin; C048496:N(6)-carboxymethyllysine; D008239:Lysine