Glutathionylation of Adenine Nucleotide Translocase Induced by Carbon Monoxide Prevents Mitochondrial Membrane Permeabilization and Apoptosis

Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal.
Journal of Biological Chemistry (Impact Factor: 4.57). 03/2010; 285(22):17077-88. DOI: 10.1074/jbc.M109.065052
Source: PubMed


The present work demonstrates the ability of CO to prevent apoptosis in a primary culture of astrocytes. For the first time, the antiapoptotic behavior can be clearly attributed to the inhibition of mitochondrial membrane permeabilization (MMP), a key event in the intrinsic apoptotic pathway. In isolated non-synaptic mitochondria, CO partially inhibits (i) loss of potential, (ii) the opening of a nonspecific pore through the inner membrane, (iii) swelling, and (iv) cytochrome c release, which are induced by calcium, diamide, or atractyloside (a ligand of ANT). CO directly modulates ANT function by enhancing ADP/ATP exchange and prevents its pore-forming activity. Additionally, CO induces reactive oxygen species (ROS) generation, and its prevention by beta-carotene decreases CO cytoprotection in intact cells as well as in isolated mitochondria, revealing the key role of ROS. On the other hand, CO induces a slight increase in mitochondrial oxidized glutathione, which is essential for apoptosis modulation by (i) delaying astrocytic apoptosis, (ii) decreasing MMP, and (iii) enhancing ADP/ATP translocation activity of ANT. Moreover, CO and GSSG trigger ANT glutathionylation, a post-translational process regulating protein function in response to redox cellular changes. In conclusion, CO protects astrocytes from apoptosis by preventing MMP, acting on ANT (glutathionylation and inhibition of its pore activity) via a preconditioning-like process mediated by ROS and GSSG.

Download full-text


Available from: Helena L A Vieira
  • Source
    • "Ischemia-reperfusion injury to the myocardium results in deglutathionylation of the Sdha subunit which decreases Complex II activity while simultaneously amplifying ROS production[52]. Adenine nucleotide translocator (ANT), which is required for ATP/ADP exchange, is also basally S-glutathionylated[53]. Decreased PGlu of ANT is associated with mitochondrial permeability transition pore (MPTP) opening and apoptosis in neurological tissue[53]. Similarly cyclophilin D is S-glutathionylated which prevents induction of mi- toptosis[54]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: At its core mitochondrial function relies on redox reactions. Electrons stripped from nutrients are used to form NADH and NADPH, electron carriers that are similar in structure but support different functions. NADH supports ATP production but also generates reactive oxygen species (ROS), superoxide (O2·-) and hydrogen peroxide (H2O2). NADH-driven ROS production is counterbalanced by NADPH which maintains antioxidants in an active state. Mitochondria rely on a redox buffering network composed of reduced glutathione (GSH) and peroxiredoxins (Prx) to quench ROS generated by nutrient metabolism. As H2O2 is quenched, NADPH is expended to reactivate antioxidant networks and reset the redox environment. Thus, the mitochondrial redox environment is in a constant state of flux reflecting changes in nutrient and ROS metabolism. Changes in redox environment can modulate protein function through oxidation of protein cysteine thiols. Typically cysteine oxidation is considered to be mediated by H2O2 which oxidizes protein thiols (SH) forming sulfenic acid (SOH). However, problems begin to emerge when one critically evaluates the regulatory function of SOH. Indeed SOH formation is slow, non-specific, and once formed SOH reacts rapidly with a variety of molecules. By contrast, protein S-glutathionylation (PGlu) reactions involve the conjugation and removal of glutathione moieties from modifiable cysteine residues. PGlu reactions are driven by fluctuations in the availability of GSH and oxidized glutathione (GSSG) and thus should be exquisitely sensitive to changes ROS flux due to shifts in the glutathione pool in response to varying H2O2 availability. Here, we propose that energy metabolism-linked redox signals originating from mitochondria are mediated indirectly by H2O2 through the GSH redox buffering network in and outside mitochondria. This proposal is based on several observations that have shown that unlike other redox modifications PGlu reactions fulfill the requisite criteria to serve as an effective posttranslational modification that controls protein function.
    Full-text · Article · Aug 2016
  • Source
    • "In the Central Nervous System (CNS) CO limits neuroinflammation [29] [30] and promotes cytoprotection in in vivo cerebral ischemia [11] [31]. In vitro experiments showed that CO prevents apoptosis in neurons [32] [33] and in astrocytes [34] [35]. Astrocytes play very important roles in the brain, conferring physical and metabolic support to neurons, releasing gliotransmitters and up-taking toxic factors. "

    Full-text · Dataset · Aug 2015
  • Source
    • "By using isolated mitochondria techniques, it was possible to demonstrate that CO directly targets mitochondria preventing MMP induced by calcium and atractyloside treatment. In nonsynaptic mitochondria isolated from brain cortex (Queiroga et al., 2010) and from mitochondria isolated liver (Queiroga et al., 2011), low amounts of CO gas inhibit several processes related to MMP: mitochondrial swelling, loss of mitochondrial membrane potential, permeabilization of inner membrane to molecules lower then 800 Da and the release of cytochrome c. Similarly to intact cells, in isolated mitochondria, ROS generation is crucial for CO to regulate MMP, since CO protection is reverted in the presence of β-carotene (Queiroga et al., 2010, 2011). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Carbon monoxide (CO) is an endogenously produced gasotransmitter, which is associated with cytoprotection and cellular homeostasis in several distinct cell types and tissues. CO mainly targets mitochondria because: (i) mitochondrial heme-proteins are the main potential candidates for CO to bind, (ii) many CO's biological actions are dependent on mitochondrial ROS signaling and (iii) heme is generated in the mitochondrial compartment. Mitochondria are the key cell energy factory, producing ATP through oxidative phosphorylation and regulating cell metabolism. These organelles are also implicated in many cell signaling pathways and the production of reactive oxygen species (ROS). Finally, mitochondria contain several factors activating programmed cell death pathways, which are released from the mitochondrial inter-membrane space upon mitochondrial membrane permeabilization. Therefore, disclosing CO mode of action at mitochondria opens avenues for deeper understanding CO's biological properties. Herein, it is discussed how CO affects the three main aspects of mitochondrial modulation of cell function: metabolism, redox response and cell death.
    Full-text · Article · Feb 2015 · Frontiers in Physiology
Show more