{"doi":"10.1073/pnas.2417572121","title":"Proteome-wide bioinformatic annotation and functional validation of the monotopic phosphoglycosyl transferase superfamily","abstract":"Phosphoglycosyl transferases (PGTs) are membrane proteins that initiate glycoconjugate biosynthesis by transferring a phospho-sugar moiety from a soluble nucleoside diphosphate sugar to a membrane-embedded polyprenol phosphate acceptor. The centrality of PGTs in complex glycan assembly and the current lack of functional information make these enzymes high-value targets for biochemical investigation. In particular, the small monotopic PGT family is exclusively bacterial and represents the minimal functional unit of the monotopic PGT superfamily. Here, we combine a sequence similarity network analysis with a generalizable, luminescence-based activity assay to probe the substrate specificity of this family of monoPGTs in the bacterial cell-membrane fraction. This strategy allows us to identify specificity on a far more significant scale than previously achievable and correlate preferred substrate specificities with predicted structural differences within the conserved monoPGT fold. Finally, we present the proof-of-concept for a small-scale inhibitor screen (eight nucleoside analogs) with four monoPGTs of diverse substrate specificity, thus building a foundation for future inhibitor discovery initiatives.","journal":"Proceedings of the National Academy of Sciences","year":2024,"id":466804,"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":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9511,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1064777,"name":"Greg J. Dodge","orcid":"0000-0002-6555-8350","position":1,"is_corresponding":false},{"id":1298924,"name":"Roxanne P. Siuda","orcid":"0000-0001-7387-8429","position":2,"is_corresponding":false},{"id":999940,"name":"Hugh Higinbotham","orcid":"0000-0002-9418-7958","position":3,"is_corresponding":false},{"id":400627,"name":"Christine A. Arbour","orcid":"0000-0001-6056-296X","position":4,"is_corresponding":false},{"id":274577,"name":"Soumi Ghosh","orcid":"0000-0001-6101-0147","position":5,"is_corresponding":false},{"id":454927,"name":"Karen N. Allen","orcid":"0000-0001-7296-0551","position":6,"is_corresponding":false},{"id":400628,"name":"Barbara Imperiali","orcid":"0000-0002-5749-7869","position":7,"is_corresponding":false},{"id":1298923,"name":"Theo Durand","orcid":"0009-0004-8131-581X","position":0,"is_corresponding":true}],"reference_count":64,"raw_metadata":null,"created_at":"2026-07-19T02:05:06.743102Z","pmid":"39602253","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":[]}