title: An aggregate-prone mutant of human glyceraldehyde-3-phosphate dehydrogenase augments oxidative stress-induced cell death in SH-SY5Y cells.
abstract: Glycerladehyde-3-phosphate dehydrogenase (GAPDH), a classic glycolytic enzyme, also has a role in mediating cell death under oxidative stress. Our previous reports suggest that oxidative stress-induced GAPDH aggregate formation is, at least in part, a mechanism to account for the death signaling. Here we show that substitution of cysteine for serine-284 of human GAPDH (S284C-GAPDH) leads to aggregate-prone GAPDH, and that its expression in SH-SY5Y human neuroblastoma results in greater dopamine-induced cell death than expression of wild type-GAPDH. Treatment of purified recombinant S284C-GAPDH in vitro with the nitric oxide donor NOR3 led to greater aggregation than wild type-GAPDH. Several lines of structural analysis revealed that S284C-GAPDH was amyloidogenic. Overexpression of doxycycline-inducible S284C-GAPDH in SH-SY5Y cells accelerated dopamine treatment-induced death and increased formation of GAPDH aggregates, compared to cells expressing wild type-GAPDH. These results suggest that aggregate-prone mutations of GAPDH such as S284C-GAPDH may confer risk of oxidative stress-induced cell death.
authors.affiliation: Laboratory of Veterinary Pharmacology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 5988531, Japan. hnakajima@vet.osakafu-u.ac.jp
authors.forename: Hidemitsu
authors.initials: H
authors.lastname: Nakajima
authors.forename: Wataru
authors.initials: W
authors.lastname: Amano
authors.forename: Ayano
authors.initials: A
authors.lastname: Fukuhara
authors.forename: Takeya
authors.initials: T
authors.lastname: Kubo
authors.forename: Shouhei
authors.initials: S
authors.lastname: Misaki
authors.forename: Yasu-Taka
authors.initials: YT
authors.lastname: Azuma
authors.forename: Takashi
authors.initials: T
authors.lastname: Inui
authors.forename: Tadayoshi
authors.initials: T
authors.lastname: Takeuchi
mesh_terms: D017209:Apoptosis; D002460:Cell Line; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006898:Hydroxylamines / Q000494:pharmacology; D009154:Mutation; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009574:Nitro Compounds; D018384:Oxidative Stress / Q000235:genetics*
chemical_list: D006898:Hydroxylamines; D020030:Nitric Oxide Donors; D009574:Nitro Compounds; C061872:FK 409; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases