My research is focused on investigating the function of the two major classes of cytosolic sHSPs, class I and II, in Arabidopsis thaliana. I created transgenic plants with increased (constitutive) or decreased (by RNAi) expression levels of either Class I or Class II cytosolic sHSPs. I also generated plants expressing affinity-tagged versions of these two sHSPs, regulated either by the CaMV 35S promoter or the sHSP’s own, heat-inducible promoter. I isolated and identified proteins that showed heat-specific association with either the Class I or Class II sHSPs. I am also studying how sHSPs recognize substrate in the model genetic organism Synechocystis sp. PCC 6803. To achieve that I use wild type or mutant sHSPs to determine how N-terminal arm mutations impact native substrate binding and processing in vivo.
I am also interested in studying the role of sHSP dynamics in substrate binding using NMR. To achieve that I am producing uniformly labeled 2H sample of TaHsp16.9 and PsHsp18.1 prepared with NMR active 13CH3 labels at Ile-delta1, Val gamma1/gamma2 and Leu delta1/delta2. I am hoping also to study the dynamic information on the substrate by using labeled substrate in the presence of unlabeled sHSP.