biostudies-literatureHuang WNIAID NIH HHSNational Institutes of HealthNIH HHS8967-8978https://www.ebi.ac.uk/biostudies/studies/S-EPMC11347161biostudies-literatureUnknown52(15)The Proline-rich Antimicrobial Peptide (PrAMP) apidaecin (Api) inhibits translation by binding in the ribosomal nascent peptide exit tunnel, trapping release factors RF1 or RF2, and arresting ribosomes at stop codons. To explore the extent of sequence variations of the native 18-amino acid Api that allows it to preserve its activity, we screened a library of synthetic mutant Api genes expressed in bacterial cells, resulting in nearly 350000 peptide variants with multiple substitutions. By applying orthogonal negative and positive selection strategies, we identified a number of multi-substituted Api variants capable of arresting ribosomes at stop codons. Our findings underscore the critical contribution of specific amino acid residues of the peptide for its on-target function while significNucleic acids researchSequence diversity of apidaecin-like peptides arresting the terminating ribosome.PMC11347161R01 AI162961Baliga CVazquez-Laslop NHuang WMankin ASfalseSequence diversity of apidaecin-like peptides arresting the terminating ribosome.The Proline-rich Antimicrobial Peptide (PrAMP) apidaecin (Api) inhibits translation by binding in the ribosomal nascent peptide exit tunnel, trapping release factors RF1 or RF2, and arresting ribosomes at stop codons. To explore the extent of sequence variations of the native 18-amino acid Api that allows it to preserve its activity, we screened a library of synthetic mutant Api genes expressed in bacterial cells, resulting in nearly 350000 peptide variants with multiple substitutions. By applying orthogonal negative and positive selection strategies, we identified a number of multi-substituted Api variants capable of arresting ribosomes at stop codons. Our findings underscore the critical contribution of specific amino acid residues of the peptide for its on-target function while signific2024-01-01T00:00:00Z2024 Aug2026-04-22T03:14:37.995Z2024-11-20T06:17:06.238ZS-EPMC113471613895315910.1093/nar/gkae567