Abstract
Publisher Summary This chapter discusses the measurement of glutathione, glutathione disulfide, and other thiols in mammalian cell and tissue homogenates using high-performance liquid chromatography (HPLC) separation of N-(1-Pyrenyl)maleimide (NPM) derivatives. As glutathione and cellular thiol-related research progresses, the role of these thiols has been extended from acting only in the maintenance of steady-state redox equilibrium to a proposed involvement in many stress induced metabolic and bioregulator functions, including signal transduction and gene expression. The NPM assay provides a rapid and simple method for analyzing both oxidized and reduced glutathione, as well as other thiols, including cysteine, 7-glutamylcysteine, homocysteine, cysteinylglycine, and N-acetylcysteine. The NPM assay provides an excellent method for determining modulations in intracellular thiols caused by oxidative stress. Because changes in redox potential have been associated with alterations in metabolism, signal transduction, and gene expression, the NPM assay provides a sensitive and accurate means of correlating thiol status with these biological processes.
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