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Preparation of Semisynthetic Peptide Macrocycles Using Split Inteins

Part of the Methods in Molecular Biology book series (MIMB,volume 1495)

Abstract

Cyclic peptides are highly desired molecules not only for basic research but also for many biomedical and pharmacological applications. Due to their potentially superior physicochemical properties as compared to their linear counterparts, they are considered as ideal candidates for studying protein–protein interactions, among others. Most of the methods developed in recent years to prepare cyclic peptides focus either on a synthetic or a recombinant route. While the former provides access to diversified, noncanonical peptides, including unnatural and d-amino acid, for example, the latter can harness the power of genetic randomization to generate and select from large peptide libraries. Only few approaches have been reported to prepare semisynthetic macrocycles that would benefit from both the advantages associated with synthetic and genetically encoded parts. We describe in this chapter a chemo-enzymatic method to make semisynthetic cyclic peptides in vitro from two fragments using protein trans-splicing and bioorthogonal oxime ligation.

Keywords

  • Antibiotics
  • Cyclic peptide
  • Intein
  • Oxime ligation
  • Protein trans-splicing
  • Ssp DnaB intein

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Acknowledgements

We thank Peter G. Schultz (Scripps Research Institute, La Jolla) for providing the plasmid for AcF incorporation. We acknowledge financial support from the DFG (MO1073/3-2, SPP1623 and Cells in Motion cluster EXC1003) and the International Graduate School of Chemistry in Münster (GSC-MS; Ph.D. stipend to S. P.).

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Correspondence to Henning D. Mootz .

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Palei, S., Mootz, H.D. (2017). Preparation of Semisynthetic Peptide Macrocycles Using Split Inteins. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_6

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  • DOI: https://doi.org/10.1007/978-1-4939-6451-2_6

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  • Publisher Name: Humana Press, New York, NY

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