The aim of this project is to characterize functional relationships for the amino acids that can affect substrate specificity as well as protein-protein interactions in glutathione S-transferases from the Thai malaria vector Anopheles dirus. We will obtain new GSTs from different classes as well as enzyme sequence variants generated by site-directed mutagenesis for kinetic characterization studies. The available crystal structures in my laboratory and these new protein forms will yield data that will increase our understanding of the mechanism of GSTs in their role in resistance to insecticides. We will also continue studying the protein-protein interactions of the GSTs and the Jun N-terminal kinase (JNK) pathway proteins. This will provide information on other physiological functions of GSTs and will increase our understanding of the interaction and regulation of proteins in vivo. This GST-JNK interaction is important as the JNK pathway has been shown to be involved in cell growth, cell differentiation, the cell stress response and apoptosis or cell death.
To achieve this goal the project entails continuing several aspects of the research previously funded by the Thailand Research Fund. These aspects include the following:
1 To continue to obtain new recombinant GST enzymes for kinetic characterization studies including enzyme sequence variants generated by site-directed mutagenesis.
2 To continue to obtain more GST crystal structures and to use the now available An. dirus GST crystal structures to provide a basis for understanding the mechanism of GSTs in their role in resistance to insecticides.
3 To continue studying the protein-protein interactions of the GSTs and the JNK kinase pathway proteins.
