title: Glyceraldehyde-3-phosphate dehydrogenase aggregate formation participates in oxidative stress-induced cell death.
abstract: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)(2) is a classic glycolytic enzyme that also mediates cell death by its nuclear translocation under oxidative stress. Meanwhile, we previously presented that oxidative stress induced disulfide-bonded GAPDH aggregation in vitro. Here, we propose that GAPDH aggregate formation might participate in oxidative stress-induced cell death both in vitro and in vivo. We show that human GAPDH amyloid-like aggregate formation depends on the active site cysteine-152 (Cys-152) in vitro. In SH-SY5Y neuroblastoma, treatment with dopamine decreases the cell viability concentration-dependently (IC(50) = 202 microM). Low concentrations of dopamine (50-100 microM) mainly cause nuclear translocation of GAPDH, whereas the levels of GAPDH aggregates correlate with high concentrations of dopamine (200-300 microM)-induced cell death. Doxycycline-inducible overexpression of wild-type GAPDH in SH-SY5Y, but not the Cys-152-substituted mutant (C152A-GAPDH), accelerates cell death accompanying both endogenous and exogenous GAPDH aggregate formation in response to high concentrations of dopamine. Deprenyl, a blocker of GAPDH nuclear translocation, fails to inhibit the aggregation both in vitro and in cells but reduced cell death in SH-SY5Y treated with only a low concentration of dopamine (100 microM). These results suggest that GAPDH participates in oxidative stress-induced cell death via an alternative mechanism in which aggregation but not nuclear translocation of GAPDH plays a role. Moreover, we observe endogenous GAPDH aggregate formation in nigra-striatum dopaminergic neurons after methamphetamine treatment in mice. In transgenic mice overexpressing wild-type GAPDH, increased dopaminergic neuron loss and GAPDH aggregate formation are observed. These data suggest a critical role of GAPDH aggregates in oxidative stress-induced brain damage.
authors.affiliation: Laboratory of Veterinary Pharmacology, Graduate School of Science, Osaka Prefecture University, Izumisano 5988531, Osaka, Japan. hnakajima@vet.oskafu-u.ac.jp
authors.forename: Hidemitsu
authors.initials: H
authors.lastname: Nakajima
authors.forename: Wataru
authors.initials: W
authors.lastname: Amano
authors.forename: Takeya
authors.initials: T
authors.lastname: Kubo
authors.forename: Ayano
authors.initials: A
authors.lastname: Fukuhara
authors.forename: Hideshi
authors.initials: H
authors.lastname: Ihara
authors.forename: Yasu-Taka
authors.initials: YT
authors.lastname: Azuma
authors.forename: Hisao
authors.initials: H
authors.lastname: Tajima
authors.forename: Takashi
authors.initials: T
authors.lastname: Inui
authors.forename: Akira
authors.initials: A
authors.lastname: Sawa
authors.forename: Tadayoshi
authors.initials: T
authors.lastname: Takeuchi
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D020134:Catalytic Domain; D016923:Cell Death; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D003545:Cysteine / Q000737:chemistry; D004298:Dopamine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D006019:Glycolysis; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D018384:Oxidative Stress
chemical_list: D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D003545:Cysteine; D004298:Dopamine