Poster Presentation 9th General Meeting of the International Proteolysis Society 2015

Regulation of cathepsin D through a propeptide-binding exosite  (#146)

Iva Hanova 1 , Jiri Brynda 1 , Radka Hobizalova 1 , Daniel Sojka 2 , Petr Kopacek 2 , Martin Horn 1 , Michael Mares 1
  1. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic
  2. Institute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, Ceske Budejovice, Czech Republic

Cathepsin D-type aspartic proteases are importantly involved in acidic proteolysis in vertebrates and invertebrates. They are biosynthesized as inactive zymogens in which the propeptide sterically blocks the active site. We investigated the mechanism of the propeptide interaction in IrCD1, a hemoglobinolytic cathepsin D of the hard tick Ixodes ricinus. First, we determined crystal structures of the IrCD1 zymogen and the mature enzyme generated by autocatalytic removal of the propeptide. An exosite was identified in the enzyme core that plays a critical role in binding the propeptide. Second, we showed that the synthetic exosite-binding segment of the propeptide is an effective inhibitor of the mature IrCD1. Its interaction associated with induced conformational changes was demonstrated by the crystal structure of the complex. The study provides the first structure of cathepsin D zymogen and defines the propeptide-binding exosite as a target for the development of specific cathepsin D inhibitors.