Protein S-glutathiolation, the reversible covalent addition of glutathione to cysteine residues on target proteins, is emerging as a candidate mechanism by which both changes in the intracellular redox state and the generation of reactive oxygen and nitrogen species may be transduced into a functional response. This review will provide an introduction to the concepts of oxidative and nitrosative stress and outline the molecular mechanisms of protein regulation by oxidative and nitrosative thiol-group modifications. Special attention will be paid to recently published work supporting a role for S-glutathiolation in stress signalling pathways and in the adaptive cellular response to oxidative and nitrosative stress. Finally, novel insights into the molecular mechanisms of S-glutathiolation as well as methodological problems related to the interpretation of the biological relevance of this post-translational protein modification will be discussed.Keywords: cysteine; glutathione; nitric oxide; reactive nitrogen species; reactive oxygen species.The regulation of protein structure and function governs the adaptive response of a cell to a changing environment. Wellestablished mechanisms for such regulation include protein± protein interactions, allosteric changes induced by the binding of ligands, proteolytic processing, and chemical modifications such as acylation, acetylation, alkylation and phosphorylation. However, the molecular mechanisms by which a cell senses the generation of reactive oxygen and nitrogen species (ROS and RNS, respectively) are still poorly understood. The aim of this review is to give an overview of the regulation of protein function by ROS/RNS, focusing on recent developments in the field that suggest that S-glutathiolation, i.e. the formation of a mixed disulfide between a protein thiol and the tripeptide glutathione (GSH), is a potential ROS/RNS-sensing mechanism which may serve to transduce oxidative and nitrosative stimuli into a functional response at various levels of cellular signalling.
The concepts of oxidative and nitrosative stressThe turnover of oxygen by cells of aerobic organisms is associated with the generation of ROS, including singlet oxygen, H 2 O 2 , superoxide and hydroxyl radicals [1]. In addition, ROS are produced by the cell as signalling molecules during cell differentiation, cell cycle progression, and in response to extracellular stimuli, including peroxisome proliferators, growth factors and mitogens [2±6]. The dark side of oxygen biochemistry [7,8] refers to the potentially hazardous effects of ROS, such as extensive oxidative damage to membrane lipids [9], DNA [10] and proteins [11] and peroxidation of lipoproteins [12]. To cope with oxidative insult, the cell has developed a number of defence strategies, both at the level of oxidative damage repair and ROS scavenging mechanisms [13±15]. Repair mechanisms that confer resistance to oxidative stress include various protein disulfide reductase enzymes as well as multifunctional DNA repair and thiol-reducing proteins ...