HV-BBI--a novel amphibian skin Bowman-Birk-like trypsin inhibitor.

Here we describe the isolation of a novel C-terminally amidated octadecapeptide--SVIGCWTKSIPPRPCFVK-amide--that contains a disulphide loop between Cys(5) and Cys(15) that is consistent with a Bowman-Birk type protease inhibitor, from the skin secretion of the Chinese Bamboo odorous frog, Huia versabilis. Named HV-BBI, the peptide is encoded by a single precursor of 62 amino acid residues whose primary structure was deduced from cloned skin cDNA. The precursor exhibits the typical organization of that encoding an amphibian skin peptide with a highly-conserved signal peptide, an intervening acidic amino acid residue-rich domain and a single HV-BBI-encoding domain located towards the C-terminus. A synthetic replicate of HV-BBI, with the wild-type K (Lys-8) residue in the presumed P1 position, was found to be a potent inhibitor of trypsin with a K(i) just slightly less than 19 nM. Substitution at this site with R (Arg) resulted in a significant reduction in potency (K(i) 57 nM), whereas replacement of K with F (Phe) resulted in the complete abolition of trypsin inhibitory activity. Thus, HV-BBI is a potent inhibitor of trypsin and the lysyl (K) residue that occupies the P1 position appears to be optimal for potency of action against this protease.