{"doi":"10.1021/jacs.0c04675","title":"Bifunctional Nitrone-Conjugated Secondary Metabolite Targeting the Ribosome","abstract":"Many microorganisms possess the capacity for producing multiple antibiotic secondary metabolites. In a few notable cases, combinations of secondary metabolites produced by the same organism are used in important combination therapies for treatment of drug-resistant bacterial infections. However, examples of conjoined roles of bioactive metabolites produced by the same organism remain uncommon. During our genetic functional analysis of oxidase-encoding genes in the everninomicin producer Micromonospora carbonacea var. aurantiaca, we discovered previously uncharacterized antibiotics everninomicin N and O, comprised of an everninomicin fragment conjugated to the macrolide rosamicin via a rare nitrone moiety. These metabolites were determined to be hydrolysis products of everninomicin P, a nitrone-linked conjugate likely the result of nonenzymatic condensation of the rosamicin aldehyde and the octasaccharide everninomicin F, possessing a hydroxylamino sugar moiety. Rosamicin binds the erythromycin macrolide binding site approximately 60 Å from the orthosomycin binding site of everninomicins. However, while individual ribosomal binding sites for each functional half of everninomicin P are too distant for bidentate binding, ligand displacement studies demonstrated that everninomicin P competes with rosamicin for ribosomal binding. Chemical protection studies and structural analysis of everninomicin P revealed that everninomicin P occupies both the macrolide- and orthosomycin-binding sites on the 70S ribosome. Moreover, resistance mutations within each binding site were overcome by the inhibition of the opposite functional antibiotic moiety binding site. These data together demonstrate a strategy for coupling orthogonal antibiotic pharmacophores, a surprising tolerance for substantial covalent modification of each antibiotic, and a potential beneficial strategy to combat antibiotic resistance.","journal":"Journal of the American Chemical Society","year":2020,"id":88351,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":10,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9408,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":447756,"name":"Michael Graf","orcid":"0000-0002-4879-3834","position":1,"is_corresponding":false},{"id":436684,"name":"Dagmara K. Derewacz","orcid":null,"position":2,"is_corresponding":false},{"id":448548,"name":"Fabian Nguyen","orcid":null,"position":3,"is_corresponding":false},{"id":232770,"name":"Jeffrey M. Spraggins","orcid":"0000-0001-9198-5498","position":4,"is_corresponding":false},{"id":447757,"name":"Maximiliane Wieland","orcid":"0000-0002-6261-0117","position":5,"is_corresponding":false},{"id":435382,"name":"Audrey E. Yñigez‐Gutierrez","orcid":"0000-0002-8215-8586","position":6,"is_corresponding":false},{"id":447758,"name":"Benjamin J. Reisman","orcid":"0000-0001-7847-2612","position":7,"is_corresponding":false},{"id":225953,"name":"Boris Zinshteyn","orcid":"0000-0003-0103-3501","position":8,"is_corresponding":false},{"id":435384,"name":"K.M. McCulloch","orcid":"0000-0001-9077-3821","position":9,"is_corresponding":false},{"id":315280,"name":"T.M. Iverson","orcid":"0000-0001-8816-6352","position":10,"is_corresponding":false},{"id":17827,"name":"Rachel Green","orcid":"0000-0001-9337-2003","position":11,"is_corresponding":false},{"id":17892,"name":"Daniel N. Wilson","orcid":"0000-0003-3816-3828","position":12,"is_corresponding":false},{"id":435385,"name":"Brian O. Bachmann","orcid":"0000-0001-5815-1176","position":13,"is_corresponding":false},{"id":435381,"name":"Emilianne M. Limbrick","orcid":"0000-0001-5020-1917","position":0,"is_corresponding":true}],"reference_count":31,"raw_metadata":null,"created_at":"2026-07-18T22:00:56.736935Z","pmid":"32709196","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}