CRISPR-based oligo recombineering prioritizes apicomplexan cysteines for drug discovery.

Autor: Benns HJ; Department of Life Sciences, Imperial College London, London, UK.; Department of Chemistry, Imperial College London, London, UK., Storch M; London Biofoundry, Imperial College Translation & Innovation Hub, London, UK., Falco JA; Department of Chemistry, Boston College, Boston, MA, USA., Fisher FR; Department of Life Sciences, Imperial College London, London, UK., Tamaki F; Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, UK., Alves E; Department of Life Sciences, Imperial College London, London, UK., Wincott CJ; Department of Life Sciences, Imperial College London, London, UK., Milne R; Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, UK., Wiedemar N; Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, UK., Craven G; Department of Chemistry, Imperial College London, London, UK., Baragaña B; Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, UK., Wyllie S; Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, UK., Baum J; Department of Life Sciences, Imperial College London, London, UK.; School of Biomedical Sciences, UNSW, Sydney, NSW, Australia., Baldwin GS; Department of Life Sciences, Imperial College London, London, UK., Weerapana E; Department of Chemistry, Boston College, Boston, MA, USA., Tate EW; Department of Chemistry, Imperial College London, London, UK. e.tate@imperial.ac.uk., Child MA; Department of Life Sciences, Imperial College London, London, UK. m.child@imperial.ac.uk.
Jazyk: angličtina
Zdroj: Nature microbiology [Nat Microbiol] 2022 Nov; Vol. 7 (11), pp. 1891-1905. Date of Electronic Publication: 2022 Oct 20.
DOI: 10.1038/s41564-022-01249-y
Abstrakt: Nucleophilic amino acids are important in covalent drug development yet underutilized as anti-microbial targets. Chemoproteomic technologies have been developed to mine chemically accessible residues via their intrinsic reactivity towards electrophilic probes but cannot discern which chemically reactive sites contribute to protein function and should therefore be prioritized for drug discovery. To address this, we have developed a CRISPR-based oligo recombineering (CORe) platform to support the rapid identification, functional prioritization and rational targeting of chemically reactive sites in haploid systems. Our approach couples protein sequence and function with biological fitness of live cells. Here we profile the electrophile sensitivity of proteinogenic cysteines in the eukaryotic pathogen Toxoplasma gondii and prioritize functional sites using CORe. Electrophile-sensitive cysteines decorating the ribosome were found to be critical for parasite growth, with target-based screening identifying a parasite-selective anti-malarial lead molecule and validating the apicomplexan translation machinery as a target for ongoing covalent ligand development.
(© 2022. The Author(s).)
Databáze: MEDLINE
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