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Vit B6 2.Transmethylation reaction 3.S-adenosylhomocysteine hydrolase 5.gama-cystathionase 4.cystathionine -synthase 6,-glutamylcysteine synthetase (GCL) 7.GSH synthetase Glutamate-cysteine ligase (GCL) activity Other major determinant of the rate of GSH synthesis Affecting heavy (GCLC) or both the heavy and light (GCLM) Transcriptional and post-transcriptional regulation of both subunits Post-transcriptional -both mRNA stabilization -destabilization and -post-translational modification Activity of GCL will be influenced by : Oxidative stress, insulin and many others increase GCL transcription or activity in variety of cells Dietary protein deficiency,dexamethasone, and GCL phosphorylation decrease GCS transcription or activity NO production NO production loss of GSH NO production prevent GSH depletion Glutathione synthase (GS) -L-glutamyl-L-cysteine + glycine + ATP GSH + ADP + Pi The regions of the active site that bind glycine and the cysteinyl moiety of -glutamylcysteine are highly specific Its activity is modified ADP GSH synthase is not subject to feedback inhibition by GSH GSH synthase deficiency -glutamylcysteine is converted to 5-oxoproline g-Glu-Cys-Gly S S g-Glu-Cys-Gly Biosynthesis of glutathione GSH synthase (GS) Glutamate-cysteine ligase (GCL) disulfide bond Glutathione distribution Glutathione synthesis in the body occure mainly in liver About 80 percent of the GSH synthesized in the liver is exported from the hepatocytes Most of this is utilized by the kidneys GSH, to supply cysteine as needed Circulating GSH is safe