{"doi":"10.1016/j.jbc.2024.107944","title":"N-glycan core tri-fucosylation requires Golgi α-mannosidase III activity that impacts nematode growth and behavior","abstract":"N-glycans with complex core chitobiose modifications are observed in various free-living and parasitic nematodes but are absent in mammals. Using Caenorhabditis elegans as a model, we demonstrated that the core N-acetylglucosamine (GlcNAc) residues are modified by three fucosyltransferases (FUTs), namely FUT-1, FUT-6, and FUT-8. Interestingly, FUT-6 can only fucosylate N-glycans lacking the α1,6-mannose upper arm, indicating that a specific α-mannosidase is required to generate substrates for subsequent FUT-6 activity. By analyzing the N-glycomes of aman-3 KOs using offline HPLC-MALDI-TOF MS/MS, we observed that the absence of aman-3 abolishes α1,3-fucosylation of the distal GlcNAc of N-glycans, which suggests that AMAN-3 is the relevant mannosidase on whose action FUT-6 depends. Enzymatic characterization of recombinant AMAN-3 and confocal microscopy studies using a knock-in strain (aman-3::eGFP) demonstrated a Golgi localization. In contrast to the classical Golgi α-mannosidase II (AMAN-2), AMAN-3 displayed a cobalt-dependent α1,6-mannosidase activity toward N-glycans. Using AMAN-3 and other C. elegans glycoenzymes, we were able to mimic nematode N-glycan biosynthesis in vitro by remodeling a fluorescein conjugated-glycan and generate a tri-fucosylated structure. In addition, using a high-content computer-assisted C. elegans analysis platform, we observed that aman-3 deficient worms display significant developmental delays, morphological, and behavioral alterations in comparison to the WT. Our data demonstrated that AMAN-3 is a Golgi α-mannosidase required for core fucosylation of the distal GlcNAc of N-glycans. This enzyme is essential for the formation of the unusual tri-fucosylated chitobiose modifications in nematodes, which may play important roles in nematode development and behavior.","journal":"Journal of Biological Chemistry","year":2024,"id":456712,"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":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9618,"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":1282039,"name":"Florian Wӧls","orcid":null,"position":1,"is_corresponding":false},{"id":421681,"name":"Saurabh Thapliyal","orcid":"0000-0003-0363-4423","position":2,"is_corresponding":false},{"id":1281558,"name":"Elsa Arcalís","orcid":"0000-0001-9883-465X","position":3,"is_corresponding":false},{"id":1282040,"name":"Hanna Gabriel","orcid":null,"position":4,"is_corresponding":false},{"id":1282041,"name":"Sascha Kubitschek","orcid":null,"position":5,"is_corresponding":false},{"id":353374,"name":"Daniel Malzl","orcid":"0000-0001-5765-6375","position":6,"is_corresponding":false},{"id":858727,"name":"Maria Regina Strobl","orcid":"0000-0001-8247-7408","position":7,"is_corresponding":false},{"id":1281559,"name":"Dieter Palmberger","orcid":"0000-0002-2086-0396","position":8,"is_corresponding":false},{"id":1282042,"name":"Thomas Luber","orcid":null,"position":9,"is_corresponding":false},{"id":409509,"name":"Carlo Unverzagt","orcid":"0000-0001-6492-1747","position":10,"is_corresponding":false},{"id":408680,"name":"Katharina Paschinger","orcid":"0000-0002-3594-7136","position":11,"is_corresponding":false},{"id":421684,"name":"Dominique A. Glauser","orcid":"0000-0002-3228-7304","position":12,"is_corresponding":false},{"id":353376,"name":"Iain B. H. Wilson","orcid":"0000-0001-8996-1518","position":13,"is_corresponding":false},{"id":1281560,"name":"Shi Yan","orcid":"0000-0003-4949-9140","position":14,"is_corresponding":false},{"id":1281557,"name":"Jonatan Kendler","orcid":"0000-0002-2470-4868","position":0,"is_corresponding":true}],"reference_count":50,"raw_metadata":null,"created_at":"2026-07-19T02:03:32.516678Z","pmid":"39481603","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":[]}