An aggregate-prone mutant of human glyceraldehyde-3-phosphate dehydrogenase augments oxidative stress-induced cell death in SH-SY5Y cells.
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.
Nakajima
Hidemitsu
H
Laboratory of Veterinary Pharmacology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 5988531, Japan. hnakajima@vet.osakafu-u.ac.jp
Amano
Wataru
W
Fukuhara
Ayano
A
Kubo
Takeya
T
Misaki
Shouhei
S
Azuma
Yasu-Taka
YT
Inui
Takashi
T
Takeuchi
Tadayoshi
T
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*
D006898:Hydroxylamines; D020030:Nitric Oxide Donors; D009574:Nitro Compounds; C061872:FK 409; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases