In this article we describe the production and screening of a genetically encoded library of 106 lanthipeptides in Escherichia coli using the substrate-tolerant lanthipeptide synthetase ProcM. This plasmid-encoded library was combined with a bacterial reverse two-hybrid system for the interaction of the HIV p6 protein with the UEV domain of the human TSG101 protein, which is a critical protein–protein interaction for HIV budding from infected cells. Using this approach, we identified an inhibitor of this interaction from the lanthipeptide library, whose activity was verified in vitro and in cell-based virus-like particle-budding assays. Given the variety of lanthipeptide backbone scaffolds that may be produced with ProcM, this method may be used for the generation of genetically encoded libraries of natural product–like lanthipeptides containing substantial structural diversity. Such libraries may be combined with any cell-based assay to identify lanthipeptides with new biological activities.
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The authors thank S. Eyckerman for pMET7-GAG-EGFP (via Addgene, plasmid # 80605), and D. Gomez-Nicola for HEK239T cells. This work was supported by the National Institutes of Health (R37 GM 058822 to W.A.V.; F32 GM0112284 to M.C.W.), Cancer Research UK (A20185 to A.T.), and the Engineering and Physical Sciences Research Council and C4X Drug Discovery (EP/L505067/1, Ph.D. studentship for K.R.L. to A.T.), AstraZeneca (Ph.D. studentship for A.T.B. to A.T.) and the Southampton University Institute for Life Sciences (studentship for C.D. to A.T.).
The authors declare no competing interests.
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Yang, X., Lennard, K.R., He, C. et al. A lanthipeptide library used to identify a protein–protein interaction inhibitor. Nat Chem Biol 14, 375–380 (2018). https://doi.org/10.1038/s41589-018-0008-5
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