title: Oxidant-induced regulation of glutathione synthesis.
abstract: 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.
authors.affiliation: University Of Alabama At Birmingham, Alabama, USA.
authors.forename: R M
authors.initials: RM
authors.lastname: Liu
authors.forename: J
authors.initials: J
authors.lastname: Choi
authors.forename: H J
authors.initials: HJ
authors.lastname: Forman
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers; D005721:Glutamate-Cysteine Ligase / Q000235:genetics; D005978:Glutathione / Q000096:biosynthesis*; D006801:Humans; D051379:Mice; D016877:Oxidants / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D005723:gamma-Glutamyltransferase / Q000235:genetics
chemical_list: D017931:DNA Primers; D016877:Oxidants; D012333:RNA, Messenger; D005723:gamma-Glutamyltransferase; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione