Abstract
FtsH is a membrane-bound zinc-dependent metalloprotease belonging to the AAA (ATPase associated with various cellular activity) protease family. The cyanobacterium Synechocystis sp. PCC6803 (PCC6083) has four FtsH homologues. These proteases have two transmembrane helices at the N terminus, and an ATP binding domain and a protease domain at the C terminus. The FtsH encoded by slr0228 is involved in selective degradation of the light- or heat- damaged D1 protein. However, the details of the degradation pathway of the damaged D1 by FtsH are unknown.
To isolate FtsH retaining the protease activity, we constructed a mutant in which the slr0228 gene was genetically modified. A his-tag, and a Factor Xa protease recognition region which is necessary to remove the his-tag afterward, were added at the N terminus of the FtsH in the PCC6803 WT cells. In this presentation, we report purification of his-tagged FtsH from PCC6803.
