Insight on the Order of Regioselective Ultrafast Formation of Disulfide Bonds in (Antimicrobial) Peptides and Miniproteins

Disulfide-rich peptides and proteins are among the most fascinating bioactive molecules. The difficulties associated with the preparation of these targets prompted the development of various chemical strategies. Nevertheless, the production of these targets remains very challenging or elusive. Recently, we introduced a strategy for one-pot disulfide bonds formation, tackling most of the previous limitations. Nevertheless, the effect of the order of oxidation remains an underexplored issue. Here we report on the synthetic flexibility and the full rainbow of oxidation orders of three disulfide bonds in targets that lack the knot motif. In contrast, our study reveals an essential order of disulfide bond formation in the EETI-II knotted miniprotein. This synthetic strategy was straightforwardly applied for the synthesis of novel analogues of the plectasin antimicrobial peptide with enhanced activities against methicillin-resistantn Staphylococcus n aureus (MRSA) , a notorious human pathogen. .