{"doi":"10.1002/anie.202409234","title":"Gas Tunnel Engineering of Prolyl Hydroxylase Reprograms Hypoxia Signaling in Cells","abstract":"Abstract Cells have evolved intricate mechanisms for recognizing and responding to changes in oxygen (O 2 ) concentrations. Here, we have reprogrammed cellular hypoxia (low O 2 ) signaling via gas tunnel engineering of prolyl hydroxylase 2 (PHD2), a non‐heme iron dependent O 2 sensor. Using computational modeling and protein engineering techniques, we identify a gas tunnel and critical residues therein that limit the flow of O 2 to PHD2’s catalytic core. We show that systematic modification of these residues can open the constriction topology of PHD2’s gas tunnel. Using kinetic stopped‐flow measurements with NO as a surrogate diatomic gas, we demonstrate up to 3.5‐fold enhancement in its association rate to the iron center of tunnel‐engineered mutants. Our most effectively designed mutant displays 9‐fold enhanced catalytic efficiency ( k cat / K M =830±40 M −1 s −1 ) in hydroxylating a peptide mimic of hypoxia inducible transcription factor HIF‐1α, as compared to WT PHD2 ( k cat / K M =90±9 M −1 s −1 ). Furthermore, transfection of plasmids that express designed PHD2 mutants in HEK‐293T mammalian cells reveal significant reduction of HIF‐1α and downstream hypoxia response transcripts under hypoxic conditions of 1 % O 2 . Overall, these studies highlight activation of PHD2 as a new pathway to reprogram hypoxia responses and HIF signaling in cells.","journal":"Angewandte Chemie International Edition","year":2024,"id":438919,"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":0.9427,"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":1190297,"name":"Haiping Ouyang","orcid":null,"position":1,"is_corresponding":false},{"id":1190298,"name":"Joseph A. G. da Costa","orcid":null,"position":2,"is_corresponding":false},{"id":1076709,"name":"Anoop R. Damodaran","orcid":"0000-0002-2094-9956","position":3,"is_corresponding":false},{"id":382599,"name":"Yue Chen","orcid":"0000-0002-0153-9927","position":4,"is_corresponding":false},{"id":1076710,"name":"Ambika Bhagi‐Damodaran","orcid":"0000-0002-4901-074X","position":5,"is_corresponding":false},{"id":1249072,"name":"Peter Windsor","orcid":"0000-0002-9490-7046","position":0,"is_corresponding":true}],"reference_count":79,"raw_metadata":null,"created_at":"2026-07-19T02:00:43.618468Z","pmid":"39168829","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":[]}