{"doi":"10.1016/j.jbc.2023.105198","title":"Catalytic process of anhydro-N-acetylmuramic acid kinase from Pseudomonas aeruginosa","abstract":"The bacterial cell envelope is the structure with which the bacterium engages with, and is protected from, its environment. Within this envelop is a conserved peptidoglycan polymer which confers shape and strength to the cell envelop. The enzymatic processes that build, remodel, and recycle the chemical components of this cross-linked polymer are preeminent targets of antibiotics and exploratory targets for emerging antibiotic structures. We report a comprehensive kinetic and structural analysis for one such enzyme, the Pseudomonas aeruginosa anhydro-N-acetylmuramic acid (anhNAM) kinase (AnmK). AnmK is an enzyme in the peptidoglycan-recycling pathway of this pathogen. It catalyzes the pairing of hydrolytic ring opening of anhNAM with concomitant ATP-dependent phosphoryl transfer. AnmK follows a random-sequential kinetic mechanism with respect to its anhNAM and ATP substrates. Crystallographic analyses of four distinct structures (apo AnmK, AnmK:AMPPNP, AnmK:AMPPNP:anhNAM, and AnmK:ATP:anhNAM) demonstrate that both substrates enter the active site independently in an ungated conformation of the substrate subsites, with protein loops acting as gates for anhNAM binding. Catalysis occurs within a closed conformational state for the enzyme. We observe this state crystallographically using ATP-mimetic molecules. A remarkable X-ray structure for dimeric AnmK sheds light on the precatalytic and postcatalytic ternary complexes. Computational simulations in conjunction with the high-resolution X-ray structures reveal the full catalytic cycle. We further report that a P. aeruginosa strain with disrupted anmK gene is more susceptible to the β-lactam imipenem compared to the WT strain. These observations position AnmK for understanding the nexus among peptidoglycan recycling, susceptibility to antibiotics, and bacterial virulence.","journal":"Journal of Biological Chemistry","year":2023,"id":351627,"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":9,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9499,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1096640,"name":"Eva Jiménez‐Faraco","orcid":"0000-0002-5548-3252","position":1,"is_corresponding":false},{"id":739836,"name":"Rhona Feltzer","orcid":"0000-0002-2071-1323","position":2,"is_corresponding":false},{"id":439978,"name":"José M. Martín-García","orcid":"0000-0002-4558-3858","position":3,"is_corresponding":false},{"id":1096641,"name":"Bhaskara Rao Karri","orcid":"0009-0001-1008-9606","position":4,"is_corresponding":false},{"id":1096642,"name":"Balajee Ramachandran","orcid":"0000-0001-6856-0176","position":5,"is_corresponding":false},{"id":739834,"name":"Choon Kim","orcid":"0000-0003-0306-8935","position":6,"is_corresponding":false},{"id":739838,"name":"Jed F. Fisher","orcid":"0000-0002-7174-4352","position":7,"is_corresponding":false},{"id":739840,"name":"J.A. Hermoso","orcid":"0000-0002-1862-8950","position":8,"is_corresponding":false},{"id":364759,"name":"Shahriar Mobashery","orcid":"0000-0002-7695-7883","position":9,"is_corresponding":false},{"id":1096639,"name":"Amr M. El‐Araby","orcid":"0000-0002-3143-8801","position":0,"is_corresponding":true}],"reference_count":48,"raw_metadata":null,"created_at":"2026-07-19T01:12:45.897709Z","pmid":"37660917","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":[]}