Investigation of peptide splicing using two-peptide-chain analogs of trypsin inhibitor SFTI-1.

This study examines peptide splicing catalyzed by serine proteinases. A series of two-peptide-chain analogs of trypsin inhibitor SFTI-1 were designed and synthesized via the solid-phase method. All consisted of two peptide chains (also called N- and C-terminal fragments) joined together by one disulfide bridge. The analogs were incubated with bovine β-trypsin or bovine α-chymotrypsin. Analysis of MS data analysis showed that, after enzyme-catalyzed degradation of the single peptide bond between the Lys and Ser residues located at the C-terminus of the C-terminal peptide chain, a new peptide bond was formed. This bond brought together the separated peptide chains, and, as a result, monocyclic SFTI-1 was recovered. This proteolytic route of peptide rearrangement appears to be similar to peptide splicing catalyzed by proteasomes. However, the proteasome is much more complex than 'classical' serine proteinases.