Title: Rapid activation of glutamate cysteine ligase following oxidative stress.
Authors: Krejsa, Cecile M; Franklin, Christopher C; White, Collin C; Ledbetter, Jeffrey A; Schieven, Gary L; Kavanagh, Terrance J
Published In J Biol Chem, (2010 May 21)
Abstract: Glutamate cysteine ligase (GCL) catalyzes the rate-limiting step in the formation of the cellular antioxidant glutathione (GSH). The GCL holoenzyme consists of two separately coded proteins, a catalytic subunit (GCLC) and a modifier subunit (GCLM). Both GCLC and GLCM are controlled transcriptionally by a variety of cellular stimuli, including oxidative stress. This study addresses post-translational control of GCL activity, which increased rapidly in human lymphocytes following oxidative stress. Activation of GCL occurred within minutes of treatment and without any change in GCL protein levels and coincided with an increase in the proportion of GCLC in the holoenzyme form. Likewise, GCLM shifted from the monomeric form to holoenzyme and higher molecular weight species. Normal rat tissues also showed a distribution of monomeric and higher molecular weight forms. Neither GCL activation, nor the formation of holoenzyme, required a covalent intermolecular disulfide bridge between GCLC and GCLM. However, in immunoprecipitation studies, a neutralizing epitope associated with enzymatic activity was protected following cellular oxidative stress. Thus, the N-terminal portion of GCLC may undergo a change that stabilizes the GCL holoenzyme. Our results suggest that a dynamic equilibrium exists between low and high activity forms of GCL and is altered by transient oxidative stress. This provides a mechanism for the rapid post-translational activation of GCL and maintenance of cellular GSH homeostasis.
PubMed ID: 20332089
MeSH Terms: Animals; Disulfides/metabolism; Enzyme Activation/physiology; Glutamate-Cysteine Ligase/metabolism*; Glutathione/metabolism*; Holoenzymes/metabolism; Homeostasis/physiology*; Humans; Jurkat Cells; Lymphocytes/enzymology; Mice; Organ Specificity/physiology; Oxidative Stress/physiology*; Protein Structure, Tertiary; Rats; Transcription, Genetic/physiology*