Cryptic enzymatic assembly of peptides armed with β-lactone warheads

Xu, G ORCID: https://orcid.org/0000-0003-4074-5165, Torri, D ORCID: https://orcid.org/0000-0003-1297-4235, Cuesta-Hoyos, S, Panda, D, Yates, LRL ORCID: https://orcid.org/0000-0002-6432-8289, Zallot, R ORCID: https://orcid.org/0000-0002-7317-1578, Bian, K, Jia, D, Iorgu, AI, Levy, C, Shepherd, SA ORCID: https://orcid.org/0000-0002-7321-1234 and Micklefield, J ORCID: https://orcid.org/0000-0001-8951-4873 (2024) Cryptic enzymatic assembly of peptides armed with β-lactone warheads. Nature Chemical Biology, 20 (10). pp. 1371-1379. ISSN 1552-4450

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Official URL: http://dx.doi.org/10.1038/s41589-024-01657-7

Abstract

Nature has evolved biosynthetic pathways to molecules possessing reactive warheads that inspired the development of many therapeutic agents, including penicillin antibiotics. Peptides armed with electrophilic warheads have proven to be particularly effective covalent inhibitors, providing essential antimicrobial, antiviral and anticancer agents. Here we provide a full characterization of the pathways that nature deploys to assemble peptides with β-lactone warheads, which are potent proteasome inhibitors with promising anticancer activity. Warhead assembly involves a three-step cryptic methylation sequence, which is likely required to reduce unfavorable electrostatic interactions during the sterically demanding β-lactonization. Amide-bond synthetase and adenosine triphosphate (ATP)-grasp enzymes couple amino acids to the β-lactone warhead, generating the bioactive peptide products. After reconstituting the entire pathway to β-lactone peptides in vitro, we go on to deliver a diverse range of analogs through enzymatic cascade reactions. Our approach is more efficient and cleaner than the synthetic methods currently used to produce clinically important warhead-containing peptides. (Figure presented.)

Item Type:	Article (Article)
Peer-reviewed:	Yes
Date Deposited:	29 Oct 2024 09:25
Publisher:	Springer
Additional Information:	The version of record of this article, first published in Nature Chemical Biology, is available online at Publisher’s website: http://dx.doi.org/10.1038/s41589-024-01657-7
Divisions:	Faculties > Science and Engineering Faculties > Science and Engineering > Department of Life Science
Subject terms:	Humans, Lactones, Peptides, Adenosine Triphosphate, Antineoplastic Agents, Peptides, Lactones, Humans, Antineoplastic Agents, Adenosine Triphosphate, 0304 Medicinal and Biomolecular Chemistry, 0601 Biochemistry and Cell Biology, Biochemistry & Molecular Biology, 3101 Biochemistry and cell biology, 3404 Medicinal and biomolecular chemistry
Data Access Statement:	The coordinates of the CysF X-ray crystal structures were deposited to PDB 8RA0. The AlphaFold structure of BhCysFE can be accessed at https://www.alphafold.ebi.ac.uk/entry/A0A562R406 (AF-A0A562R406-F1-model_v4). Structures used for modeling and docking studies can be accessed from PDB 5BSM, 5BSR, 5WM3, 5IE3, 4FUT, 4GXR and 4GXQ. All proteins characterized in this study can be accessed from UniProt using the accession codes presented in Supplementary Tables 1–3 and their synthetic gene sequences are provided in Supplementary Data 1. The remaining data are available in the main text or the Supplementary Information. Correspondence and requests for materials should be addressed to J.M.
URI:	https://e-space.mmu.ac.uk/id/eprint/636700
DOI:	https://doi.org/10.1038/s41589-024-01657-7
ISSN	1552-4450
e-ISSN	1552-4469