Glutathione transferases in maize (zea mays)

Abstract

The glutathione transferases (GSTs) of maize have been the most studied GSTs in plants, however much is still not known about these enzymes. In the course of the current study six GST subunits (Zm GSTs I, II and III, which have been reported previously, and Zm GSTs V, VI and VII, which have not been previously reported) have been identified in the dimers Zm GST I-I, I-II, I-III, V-V, V-VI and V-VII. Maize GSTs are known to be important in herbicide detoxification and the purified maize enzymes were each found to have differing activities toward a number of herbicides, and also a range of other potential GST substrates. Additionally, Zm GST I II and Zm GST V-V possessed glutathione peroxidase activity. The developmental regulation and chemical inducibility of maize GSTs were studied in maize seedlings using western blotting, with different subunits showing markedly different responses. Zm GST I was constitutively present in all plant parts and unaffected by chemical treatment, Zm GST II was only detected in young roots but was induced in roots and shoots by many different chemical treatments, and Zm GST V was present at low levels throughout maize plants, with levels enhanced greatly by treatment with the safener dichlormid but not by other chemicals tested. cDNA clones corresponding to Zm GST subunits I, III, V, VI and VII were isolated by library screening using antibody or DNA probes. The cDNA sequences for Zm GST subunits V, VI and VH were different from those of previously cloned type I (theta class) maize GSTs and were most similar to the auxin-regulated GST family (type III or tau class GSTs) previously only identified in dicotyledonous species. The cloned GSTs were expressed as recombinant proteins in E. coli, allowing further characterisation, including detailed kinetic analysis for recombinant Zm GST I-I and Zm GST V-V.