UNIT 6.7 Oxidant-Induced Regulation of Glutathione Synthesis
Published Online: 1 AUG 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Toxicology
How to Cite
Liu, R.-M., Choi, J. and Forman, H. J. 2001. Oxidant-Induced Regulation of Glutathione Synthesis. Current Protocols in Toxicology. 8:6.7:6.7.1–6.7.21.
- Published Online: 1 AUG 2001
- Published Print: MAY 2001
This unit describes protocols for characterizing the expression of two glutathione biosynthesis enzymes: gamma-glutamylcysteine synthase (GCS) and gamma-glutamyl transpeptidase (GGT) in response to oxidants. GCS catalyzes the first and rate-limiting step of glutathione synthesis, while GGT degrades extracellular glutathione (GSH) to provide the amino acids required for intracellular synthesis of GSH. Northern blot hybridization is used to quantitatively assess the mRNAs for the two enzymes in response to oxidant. Nuclear run-on is used to determine the rate of transcription.