title: Glycation and inactivation of human Cu-Zn-superoxide dismutase. Identification of the in vitro glycated sites.
abstract: The nonenzymatic glycosylation (glycation) of Cu-Zn-superoxide dismutase led to gradual inactivation of the enzyme (Arai, K. Iizuka, S., Tada, Y., Oikawa, K., and Taniguchi, N. (1987) Biochim. Biophys. Acta 924, 292-296). The purified superoxide dismutase from human erythrocytes comprises both glycated and nonglycated forms. The nonglycated Cu-Zn-superoxide dismutase was isolated by boronate affinity chromatography. Incubation of the nonglycated superoxide dismutase with D-[6-3H]glucose in vitro resulted in the gradual accumulation of radioactivity in the enzyme protein, and Schiff base adducts were trapped by NaBH4. The sites of glycation of the superoxide dismutase were identified by amino acid analysis after reverse-phase high performance liquid chromatography of the trypsin-treated peptides. Lysine residues, i.e. Lys3, Lys9, Lys30, Lys36, Lys122, and Lys128, were found to be glycated. Three of the glycated sites lie in Lys-Gly, two in Lys-Ala, and one in Lys-Val. The inactivation of the superoxide dismutase on the glycation is due mainly to the glycation of Lys122 and Lys128, which are supposed to be located in an active site liganding loop. The remaining five sites, such as Lys-Glu, Lys-Asp, Lys-His, and Lys-Thr are relatively inactive as to the formation of either a Schiff base or an Amadori adduct.
authors: Department of Pediatric Dentistry, Hokkaido University School of Dentistry, Sapporo, Japan.
authors: K
authors: K
authors: Arai
authors: S
authors: S
authors: Maguchi
authors: S
authors: S
authors: Fujii
authors: H
authors: H
authors: Ishibashi
authors: K
authors: K
authors: Oikawa
authors: N
authors: N
authors: Taniguchi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003300:Copper / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors*; D015032:Zinc / Q000378:metabolism
chemical_list: D000596:Amino Acids; D003300:Copper; D013482:Superoxide Dismutase; D015032:Zinc