{"doi":"10.1073/pnas.2007297117","title":"Metabolic precision labeling enables selective probing of O-linked <i>N</i> -acetylgalactosamine glycosylation","abstract":"-acetylgalactosamine-4-epimerase (GALE) like conventional GalNAc-based probes. GalNAzMe enters O-GalNAc glycosylation but does not enter other major cell surface glycan types including Asn(N)-linked glycans. We transfect cells with the engineered pyrophosphorylase mut-AGX1 to biosynthesize the nucleotide-sugar donor uridine diphosphate (UDP)-GalNAzMe from a sugar-1-phosphate precursor. Tagged with a bioorthogonal azide group, GalNAzMe serves as an O-glycan-specific reporter in superresolution microscopy, chemical glycoproteomics, a genome-wide CRISPR-knockout (CRISPR-KO) screen, and imaging of intestinal organoids. Additional ectopic expression of an engineered glycosyltransferase, \"bump-and-hole\" (BH)-GalNAc-T2, boosts labeling in a programmable fashion by increasing incorporation of GalNAzMe into the cell surface glycoproteome. Alleviating the need for GALE-KO cells in metabolic labeling experiments, GalNAzMe is a precision tool that allows a detailed view into the biology of a major type of cancer-relevant protein glycosylation.","journal":"Proceedings of the National Academy of Sciences","year":2020,"id":55598,"datarank":0.6814942173405006,"base_score":4.543294782270004,"endowment":4.543294782270004,"self_citation_contribution":0.6814942173405006,"citation_network_contribution":0.0,"self_endowment_contribution":0.6814942173405006,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":93,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9619,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":288029,"name":"Ömür Y. 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Wagner","orcid":"0000-0002-8532-7496","position":8,"is_corresponding":false},{"id":288032,"name":"Ganka Bineva‐Todd","orcid":"0000-0001-8926-9417","position":9,"is_corresponding":false},{"id":288033,"name":"Aristotelis Antonopoulos","orcid":"0000-0002-1592-4092","position":10,"is_corresponding":false},{"id":288034,"name":"Anna Cioce","orcid":"0000-0001-7284-1712","position":11,"is_corresponding":false},{"id":288035,"name":"William M. Browne","orcid":"0000-0003-2647-3714","position":12,"is_corresponding":false},{"id":268920,"name":"Zhen Li","orcid":"0000-0002-8853-4941","position":13,"is_corresponding":false},{"id":288036,"name":"David C. Briggs","orcid":"0000-0002-9793-7339","position":14,"is_corresponding":false},{"id":288037,"name":"Holly Douglas","orcid":"0000-0002-2669-8852","position":15,"is_corresponding":false},{"id":62605,"name":"Gaelen T. Hess","orcid":"0000-0002-3836-0277","position":16,"is_corresponding":false},{"id":268917,"name":"Anthony J. Agbay","orcid":"0000-0003-0466-5977","position":17,"is_corresponding":false},{"id":288038,"name":"Chloë Roustan","orcid":"0000-0002-8439-8997","position":18,"is_corresponding":false},{"id":288039,"name":"Svend Kjær","orcid":"0000-0001-9767-8683","position":19,"is_corresponding":false},{"id":288040,"name":"Stuart M. Haslam","orcid":"0000-0002-5563-679X","position":20,"is_corresponding":false},{"id":288041,"name":"Ambrosius P. Snijders","orcid":"0000-0002-5416-8592","position":21,"is_corresponding":false},{"id":62612,"name":"Michael C. Bassik","orcid":"0000-0001-5185-8427","position":22,"is_corresponding":false},{"id":270551,"name":"W. E. Moerner","orcid":"0000-0002-2830-209X","position":23,"is_corresponding":false},{"id":259746,"name":"Vivian Li","orcid":"0000-0003-2041-0085","position":24,"is_corresponding":false},{"id":218284,"name":"Carolyn R. 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