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Transport to the vacuoles ATP-dependent membrane pumps recognize the glutathione S-conjugates and transfer them into the vacuoles Glutathione conjugation tags the xenobiotic substrates for recognition by membrane pumps Measurement of total glutathione S-transferase (GST) activity:spectrophotometrically at 340 nm conjugate CDNB Best model substrate: 1-chloro-2,4-dinitrobenzene (CDNB) Separation of GST isoenzymes by HPLC in wheat extracts Crude extract (NH4)2SO4 precipitation affinity chromatography on GSH-agarose column RP-HPLC UV detection (214 nm) Old history (1990s): GST isoenzymes in maize Two subunits Homo- and heterodimer molecules constitutive or inducible GSTs today Maize: 42 isoenzymes EST (expressed sequence tag) database BLAST searching (Du Pont Company, USA) Arabidopsis thaliana: 47 isoenzymes McGonigle et al., Plant Physiol

Huang T, Shu Y, Cai YD

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24 Besides, NADPH acts as the reducing reagent for 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), the rate-limiting enzyme of the mevalonate pathway, which leads to the synthesis of cholesterol and nonsterol isoprenoids