1. Introduction of Bifunctionality into the Multidomain Architecture of the ω-Ester-Containing Peptide Plesiocin.
- Author
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Lee C, Lee H, Park JU, and Kim S
- Subjects
- Biosynthetic Pathways drug effects, Chromatography, High Pressure Liquid, Chymotrypsin chemistry, Cloning, Molecular, Escherichia coli genetics, Esters chemistry, Peptides genetics, Peptides isolation & purification, Protease Inhibitors chemistry, Protease Inhibitors pharmacology, Protein Binding genetics, Protein Domains genetics, Protein Processing, Post-Translational genetics, Ribosomes chemistry, Ribosomes genetics, Trypsin chemistry, Trypsin genetics, Trypsin Inhibitors chemistry, Chymotrypsin antagonists & inhibitors, Peptides chemistry, Protein Engineering
- Abstract
The modular biosynthetic pathway of ribosomally synthesized and post-translationally modified peptides (RiPPs) enhances their engineering potential for exploring new structures and biological functions. The ω-ester-containing peptides (OEPs), a subfamily of RiPPs, have distinct side-to-side ester or amide linkages and frequently present more than one macrocyclic domain in a "beads-on-a-string" structure. In an effort to improve the engineering potential of RiPPs, we present here the idea that the multidomain architecture of an OEP, plesiocin, can be exploited to create a bifunctional modified peptide. Characterization of plesiocin variants revealed that strong chymotrypsin inhibition relies on the bicyclic structure of the domain in which a leucine residue in the hairpin loop functions as a specificity determinant. Four domains of plesiocin promote simultaneous binding of multiple enzymes, where the C-terminal domain binds chymotrypsin most efficiently. Using this information, we successfully engineered a plesiocin variant in which two different domains inhibit chymotrypsin and trypsin. This result suggests that the multidomain architecture of OEPs is a useful platform for engineering multifunctional hybrid RiPPs.
- Published
- 2020
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