Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Aggregation Causes Mitochondrial Dysfunction during Oxidative Stress-induced Cell Death

Abstract

Glycolytic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein that also mediates cell death under oxidative stress. We reported previously that the active-site cysteine (Cys-152) of GAPDH plays an essential role in oxidative stress-induced aggregation of GAPDH associated with cell death, and a C152A-GAPDH mutant rescues nitric oxide (NO)-induced cell death by interfering with the aggregation of wild type (WT)-GAPDH. However, the detailed mechanism underlying GAPDH aggregate-induced cell death remains elusive. Here we report that NO-induced GAPDH aggregation specifically causes mitochondrial dysfunction. First, we observed a correlation between NO-induced GAPDH aggregation and mitochondrial dysfunction, when GAPDH aggregation occurred at mitochondria in SH-SY5Y cells. In isolated mitochondria, aggregates of WT-GAPDH directly induced mitochondrial swelling and depolarization, whereas mixtures containing aggregates of C152A-GAPDH reduced mitochondrial dysfunction. Additionally, treatment with cyclosporin A improved WT-GAPDH aggregate-induced swelling and depolarization. In doxycycline-inducible SH-SY5Y cells, overexpression of WT-GAPDH augmented NO-induced mitochondrial dysfunction and increased mitochondrial GAPDH aggregation, whereas induced overexpression of C152A-GAPDH significantly suppressed mitochondrial impairment. Further, NO-induced cytochrome c release into the cytosol and nuclear translocation of apoptosis-inducing factor from mitochondria were both augmented in cells overexpressing WT-GAPDH but ameliorated in C152A-GAPDH-overexpressing cells. Interestingly, GAPDH aggregates induced necrotic cell death via a permeability transition pore (PTP) opening. The expression of either WT- or C152A-GAPDH did not affect other cell death pathways associated with protein aggregation, such as proteasome inhibition, gene expression induced by endoplasmic reticulum stress, or autophagy. Collectively, these results suggest that NO-induced GAPDH aggregation specifically induces mitochondrial dysfunction via PTP opening, leading to cell death.

Metabolic Reprogramming in Astrocytes Distinguishes Region-Specific Neuronal Susceptibility in Huntington Mice

“…These seemingly conflicting activities were measured in homogenates of whole brain tissues, which do not resolve affected and unaffected regions of the HD brain or the two cell-type-dependent classes of MT. Furthermore, expression of expanded polyglutamine tracts or increases in oxidative stress promotes GAPDH aggregation and catalytically inactivates the enzyme (Hwang et al, 2015; Nakajima et al, 2017). Under these conditions, cells would be expected to shift toward oxidative phosphorylation as an energy source.…”

Biological activity of peptides purified from fish skin hydrolysates

“…Involvement of GAPDH in nuclear translocation and its aggregation under oxidative stress have been proposed as processes leading to GAPDH-mediated cell death. Furthermore, oxidative stressors initiate amyloid-like GAPDH aggregation via intermolecular disulfide bonds at Cys-152 (Nakajima et al 2017). However, the antimicrobial activity observed could be as a result of the peptides, YFGAP and SJGAP, acting as analogs of GAPDH in the sixth step of glycolysis due to high similarities with its N-terminus.…”

Effects of different processing methods on the antioxidant and immune stimulating abilities of garlic

“…Oxidative stress can cause cell death (Nakajima et al., ) and can promote the activation of mediator signaling molecules such as NF‐kB to increase inflammatory cytokine production (Buelna‐Chontal & Zazueta, ). However, endogenous antioxidants such as Glutathione play critical roles in defending against oxidative stress to protect host immune cells from free radicals.…”