A new family of serine protease inhibitors (Bombyx family) as established from the unique topological relation between the positions of disulphide bridges and reactive site.

Abstract

The positions of the reactive site and the disulfide bridges in fungal protease inhibitor F (FPI-F) from silkworm (Bombyx mori), which has a unique amino acid sequence and inhibitory specificity, were investigated. At pH 3.0, subtilisin BPN', which is one of target proteases of this inhibitor, specifically cleaved the peptide bond of the inhibitor at Thr(29)-Val(30). The cleaved bond was regenerated by subtilisin BPN' at pH 8.0. These results indicate that the Thr(29)-Val(30) bond of the inhibitor is the reactive site. The locations of disulfide bridges were determined to be Cys(3)-Cys(35), Cys(14)-Cys(27), Cys(18)-Cys(55), and Cys(37)-Cys(49). Based on the positions of the reactive site and the disulfide bridges, FPI-F is considered to be a member of a new family of serine protease inhibitors. We propose the designation Bombyx family for the new inhibitor family of which FPI-F is a member.