{"doi":"10.3390/ijms22189984","title":"Identification of Small Molecule Inhibitors against Staphylococcus aureus Dihydroorotase via HTS","abstract":"Drug-resistant Staphylococcus aureus is an imminent threat to public health, increasing the importance of drug discovery utilizing unexplored bacterial pathways and enzyme targets. De novo pyrimidine biosynthesis is a specialized, highly conserved pathway implicated in both the survival and virulence of several clinically relevant pathogens. Class I dihydroorotase (DHOase) is a separate and distinct enzyme present in gram positive bacteria (i.e., S. aureus, B. anthracis) that converts carbamoyl-aspartate (Ca-asp) to dihydroorotate (DHO)—an integral step in the de novo pyrimidine biosynthesis pathway. This study sets forth a high-throughput screening (HTS) of 3000 fragment compounds by a colorimetry-based enzymatic assay as a primary screen, identifying small molecule inhibitors of S. aureus DHOase (SaDHOase), followed by hit validation with a direct binding analysis using surface plasmon resonance (SPR). Competition SPR studies of six hit compounds and eight additional analogs with the substrate Ca-asp determined the best compound to be a competitive inhibitor with a KD value of 11 µM, which is 10-fold tighter than Ca-asp. Preliminary structure–activity relationship (SAR) provides the foundation for further structure-based antimicrobial inhibitor design against S. aureus.","journal":"International Journal of Molecular Sciences","year":2021,"id":198614,"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":0.9546,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":418625,"name":"Russell P. Pesavento","orcid":"0000-0003-2599-7751","position":1,"is_corresponding":false},{"id":397429,"name":"Jinhong Ren","orcid":"0000-0003-0328-702X","position":2,"is_corresponding":false},{"id":773887,"name":"Isoo Youn","orcid":"0000-0002-8851-0720","position":3,"is_corresponding":false},{"id":624484,"name":"Youngjin Kwon","orcid":null,"position":4,"is_corresponding":false},{"id":504319,"name":"Kassapa Ellepola","orcid":"0000-0003-3806-9807","position":5,"is_corresponding":false},{"id":390591,"name":"Chun‐Tao Che","orcid":"0000-0001-7064-271X","position":6,"is_corresponding":false},{"id":397432,"name":"Michael E. Johnson","orcid":"0000-0002-8246-4081","position":7,"is_corresponding":false},{"id":352857,"name":"Hyun Lee","orcid":"0000-0003-2570-8120","position":8,"is_corresponding":false},{"id":773886,"name":"Amy J. Rice","orcid":"0000-0001-5436-4799","position":0,"is_corresponding":true}],"reference_count":23,"raw_metadata":null,"created_at":"2026-07-18T23:50:31.857233Z","pmid":"34576147","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":[]}