Sortase A-mediated synthesis of ligand-grafted cyclized peptides for modulating a model protein-protein interaction.
Specific ligand-grafted cyclic peptides are promising drug candidates that can modulate protein-protein interactions (PPIs) with increased proteolytic stability. In this study, we aimed to demonstrate that Sortase A (SrtA)-mediated peptide transpeptidation can be applied to produce bioactive sequence-grafted, stable, cyclic peptides. A naturally occurring cyclic peptide, sunflower trypsin inhibitor 1 (SFTI-1), was selected as the scaffold, and a tetrapeptide motif, Glu-Ser-Asp-Val (ESDV), was grafted into the scaffold as a model ligand. The linear precursor of the grafted peptide with SrtA-recognition motifs at the N- and C-termini was cyclized in good yield simply by co-incubation with SrtA. The ESDV-grafted cyclic SFTI-1 obtained was confirmed to have high stability against proteolysis by human serum and bound to the target PDZ2 domain of postsynaptic density-95 protein. An optimized sequence-grafted cyclic SFTI-1 could competitively suppress the interaction of PDZ2 with its natural ligand, the C-terminal peptide of the NR2B subunit of the N-methyl-D-aspartate receptor. These results show that a strategy combining peptide grafting into the SFTI-1 scaffold with SrtA-catalyzed cyclization can be a simple and effective method for producing stable peptide drugs.
Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan.
Jing
J
Zhang
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan.
Satoshi
S
Yamaguchi
Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan. nagamune@bioeng.t.u-tokyo.ac.jp.|Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan. nagamune@bioeng.t.u-tokyo.ac.jp.
Teruyuki
T
Nagamune
D019881:Aminoacyltransferases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D055808:Drug Discovery; D006801:Humans; D008958:Models, Molecular; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D055550:Protein Stability
D001426:Bacterial Proteins; D010456:Peptides, Cyclic; C448501:SFTI-1 peptide, sunflower; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
Peptide cyclization; Peptide drugs; Protein-protein interactions; Sortase A; Sunflower trypsin inhibitor 1