title: HPLC-based assays for enzymes of glutathione biosynthesis.
abstract: Glutamate cysteine ligase and glutathione synthase carry out the two-step synthesis of glutathione. The fluorescent thiol-reactive compound monobromobimane is used to derivatize reaction products in an HPLC-based assay with fluorescence detection. The assay described in this unit can be adapted for tissue homogenates or cultured cells.
authors.affiliation: University of Washington, Seattle, Washington, USA.
authors.forename: C C
authors.initials: CC
authors.lastname: White
authors.forename: C J
authors.initials: CJ
authors.lastname: Krejsa
authors.forename: D L
authors.initials: DL
authors.lastname: Eaton
authors.forename: T J
authors.initials: TJ
authors.lastname: Kavanagh
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005721:Glutamate-Cysteine Ligase / Q000378:metabolism*; D005978:Glutathione / Q000096:biosynthesis*; D005981:Glutathione Synthase / Q000378:metabolism*; D012015:Reference Standards; D013050:Spectrometry, Fluorescence
chemical_list: D005721:Glutamate-Cysteine Ligase; D005981:Glutathione Synthase; D005978:Glutathione