{"doi":"10.1002/anie.202313900","title":"The Molecular Basis of Human ALKBH3 Mediated RNA <i>N</i><sup><i>1</i></sup>‐methyladenosine (m<sup>1</sup>A) Demethylation","abstract":"Abstract N 1 ‐methyladenosine (m 1 A) is a prevalent post‐transcriptional RNA modification, and the distribution and dynamics of the modification play key epitranscriptomic roles in cell development. At present, the human AlkB Fe(II)/α‐ketoglutarate‐dependent dioxygenase family member ALKBH3 is the only known mRNA m 1 A demethylase, but its catalytic mechanism remains unclear. Here, we present the structures of ALKBH3‐oligo crosslinked complexes obtained with the assistance of a synthetic antibody crystallization chaperone. Structural and biochemical results showed that ALKBH3 utilized two β‐hairpins (β4‐loop‐β5 and β′‐loop‐β′′) and the α2 helix to facilitate single‐stranded substrate binding. Moreover, a bubble‐like region around Asp194 and a key residue inside the active pocket (Thr133) enabled specific recognition and demethylation of m 1 A‐ and 3‐methylcytidine (m 3 C)‐modified substrates. Mutation of Thr133 to the corresponding residue in the AlkB Fe(II)/α‐ketoglutarate‐dependent dioxygenase family members FTO or ALKBH5 converted ALKBH3 substrate selectivity from m 1 A to N 6 ‐methyladenosine (m 6 A), as did Asp194 deletion. Our findings provide a molecular basis for understanding the mechanisms of substrate recognition and m 1 A demethylation by ALKBH3. This study is expected to aid structure‐guided design of chemical probes for further functional studies and therapeutic applications.","journal":"Angewandte Chemie International Edition","year":2023,"id":328535,"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":23,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9596,"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":1050000,"name":"Hong‐Chao Duan","orcid":"0000-0002-5827-6021","position":1,"is_corresponding":false},{"id":513839,"name":"Marcin Paduch","orcid":"0000-0002-2223-5906","position":3,"is_corresponding":false},{"id":715239,"name":"Jingyan Hu","orcid":"0000-0003-4212-4741","position":4,"is_corresponding":false},{"id":1050001,"name":"Chi Zhang","orcid":"0000-0003-1762-0639","position":5,"is_corresponding":false},{"id":397874,"name":"Yajuan Mu","orcid":"0000-0002-1759-7291","position":6,"is_corresponding":false},{"id":716045,"name":"Hou‐Wen Lin","orcid":null,"position":7,"is_corresponding":false},{"id":21408,"name":"Chuan He","orcid":"0000-0003-4319-7424","position":8,"is_corresponding":false},{"id":260362,"name":"Anthony A. Kossiakoff","orcid":"0000-0003-3174-9359","position":9,"is_corresponding":false},{"id":1050002,"name":"Guifang Jia","orcid":"0000-0002-4186-6922","position":10,"is_corresponding":false},{"id":715246,"name":"Liang Zhang","orcid":"0000-0002-7672-1168","position":11,"is_corresponding":false},{"id":715235,"name":"Lin Zhang","orcid":"0000-0002-3076-6993","position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":null,"created_at":"2026-07-19T01:08:52.196069Z","pmid":"38158383","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":[]}