Abstract
FtsH protease is an ATP dependent metalloprotease, which localized on thylakoid membrane. Main function of FtsH protease is to degrade photo-damaged D1 protein in the reaction center of photosystem2. This protease is composed from transmembrane domain, ATPase domain and protease domain. In this study, we focused on the functional analysis of ATPase domain of FtsH protease in var2 subunit. FtsH protease from Arabidopsis was composed from at least four types of subunits, though that from E.coli was homohexamer. We constructed an expression vector of ATPase domain and transformed into BL21-CodonPlus(DE3)-RIL. Purification was performed using Ni-Chelating Sepharose. The quality of this protein was verified by western blotting and peptide sequencing. We examined biochemical analysis to determine the optimum condition of this enzyme. We also investigated effect of inhibitors. Kinetic parameters, Km,Vmax and Kcat, were calculated in the optimum condition.
