Abstract
In vitro selection methods represent a powerful approach toward identifying high-affinity peptide ligands from highly diverse peptide libraries against a desired target. We herein describe a method for the display and selection of cyclic thioether peptide libraries. Reprogramming the initiation event from fMet to an N-chloroacetyl-amino acid by utilizing flexizyme to rapidly and efficiently prepare the aa-tRNA can be effectively used to initiate translation, upon which the thiol group of an inserted cysteine at the C terminus of the designed library spontaneously reacts to yield a nonreducible cyclic thioether peptide readily compatible with any in vitro display methods. Thus, cyclic peptides already in a nonreducible stable form can be selected directly against the target of interest.
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Acknowledgments
We would like to thank Dr. Hiroshi Murakami and all of the members of the Suga lab and PeptiDream for their helpful suggestions.
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Reid, P.C., Goto, Y., Katoh, T., Suga, H. (2012). Charging of tRNAs Using Ribozymes and Selection of Cyclic Peptides Containing Thioethers. In: Douthwaite, J., Jackson, R. (eds) Ribosome Display and Related Technologies. Methods in Molecular Biology, vol 805. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-379-0_19
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DOI: https://doi.org/10.1007/978-1-61779-379-0_19
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