Glycation proceeds faster in mutated Cu, Zn‐superoxide dismutases related to familial amyotrophic lateral sclerosis

• Published in: The FASEB journal 2003-05, Vol.17 (8), p.1-18
• Main Authors: Takamiya, Rina, Takahashi, Motoko, Myint, Theingi, Park, Yong Seek, Miyazawa, Nobuko, Endo, Takeshi, Fujiwara, Noriko, Sakiyama, Haruhiko, Misonou, Yoshiko, Miyamoto, Yasuhide, Fujii, Junichi, Taniguchi, Naoyuki
• Format: Article
• Language: English
• Subjects: Amyotrophic Lateral Sclerosis - enzymology; Amyotrophic Lateral Sclerosis - genetics; Animals; Cell Line; copper; Dose-Response Relationship, Drug; Family Health; Fructose - metabolism; Glucose - metabolism; Glucose - pharmacology; Glycosylation - drug effects; Humans; hydrogen peroxide; Hydrogen Peroxide - metabolism; Mutation; Mutation, Missense; posttranslational modification; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; ROS; Spodoptera; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism
• ISSN: 0892-6638; 1530-6860
• DOI: 10.1096/fj.02-0768fje

ABSTRACT Amyotrophic lateral sclerosis (ALS) involves the progressive degeneration of motor neurons in the spinal cord and motor cortex. It has been shown that 15–20% of patients with familial ALS (FALS) have defects in the Sod1 gene that encodes Cu, Zn‐superoxide dismutase (SOD). To elucidate the pathological role of mutated Cu, Zn‐SODs in FALS, the susceptibility of mutants to glycation was examined. Mutated Cu, Zn‐SODs (G37R, G93A, and I113T) related to FALS and wild type were produced in a baculovirus/insect cell expression system. Glycated and nonglycated proteins were separated on a boronate column, and the nonglycated fraction was then incubated with glucose. The mutated Cu, Zn‐SODs were found to be highly susceptible to glycation compared with the wild‐type enzyme as estimated by Western blot analysis using an anti‐hexitol lysine antibody. The mutated Cu, Zn‐SOD incubated with glucose generated higher levels of hydrogen peroxide than the wild‐type enzyme. Mutated Cu, Zn‐SODs were also shown to be highly susceptible to fructation, and the fructated mutant also produced higher levels of hydrogen peroxide than the wild type. These results suggest that high susceptibility of mutated Cu, Zn‐SODs to glycation could be the origin of the oxidative stress associated with neuronal dysfunction in FALS.