{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T05:47:52Z","timestamp":1793339272060,"version":"4.5.0"},"reference-count":46,"publisher":"American Chemical Society (ACS)","issue":"29","license":[{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Basic Research Program of China (973 Program)","doi-asserted-by":"crossref","award":["2022YFA1304700"],"award-info":[{"award-number":["2022YFA1304700"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China (NSFC)","doi-asserted-by":"crossref","award":["22201217"],"award-info":[{"award-number":["22201217"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China (NSFC)","doi-asserted-by":"crossref","award":["32101026"],"award-info":[{"award-number":["32101026"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China (NSFC)","doi-asserted-by":"crossref","award":["32471504"],"award-info":[{"award-number":["32471504"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China (NSFC)","doi-asserted-by":"crossref","award":["92478001"],"award-info":[{"award-number":["92478001"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China (NSFC)","doi-asserted-by":"crossref","award":["T2596061"],"award-info":[{"award-number":["T2596061"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Lin Gang Laboratory","award":["LGL-2614-18"],"award-info":[{"award-number":["LGL-2614-18"]}]},{"name":"Lin Gang Laboratory","award":["LGL-2617-01"],"award-info":[{"award-number":["LGL-2617-01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,7,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Cell-surface glycans exhibit extensive microheterogeneity, yet their site-specific dynamic behavior remains largely uncharacterized due to the lack of effective analytical methods. Here, we report a chemical enrichment strategy for profiling the temporal dynamics of intact cell-surface sialylated N-glycopeptides at site-specific and single-glycoform resolution. This method integrates metabolic labeling of sialylated glycans with azide reporters, copper-free strain-promoted azide\u2013alkyne cycloaddition (SPAAC) using a photocleavable biotin probe for glycan labeling in living cells, and affinity enrichment of intact glycopeptides. Coupling this strategy with pulse\u2013chase analysis enables quantitative tracking of glycan turnover across multiple structural levels, including glycoproteins, glycosylation sites, glycans, and site-specific glycoforms. The results reveal distinct turnover kinetics among glycoforms at the same site and show that glycan-level dynamics can differ substantially from protein-level stability. This work provides a robust analytical platform for investigating cell-surface glycan dynamics with high structural resolution.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c02598","type":"journal-article","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T04:32:46Z","timestamp":1783917166000},"page":"21697-21708","source":"Crossref","is-referenced-by-count":1,"title":["Chemically\nCleavable Enrichment Enables Site-Specific\nProfiling of Cell-Surface N-Glycan Microheterogeneity Dynamics"],"prefix":"10.1021","volume":"98","author":[{"given":"Mengyao","family":"Fan","sequence":"first","affiliation":[{"name":"Chinese Academy of Sciences , , , ,","place":["Shanghai, China, 200032"]},{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengyang","family":"Ding","sequence":"additional","affiliation":[{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaoyao","family":"Liu","sequence":"additional","affiliation":[{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiheng","family":"Jiang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , , ,","place":["Shanghai, China, 200032"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiwei","family":"Zhao","sequence":"additional","affiliation":[{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"An","sequence":"additional","affiliation":[{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuyang","family":"Cao","sequence":"additional","affiliation":[{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weimin","family":"Zheng","sequence":"additional","affiliation":[{"name":"Nanjing Medical University , , , ,","place":["Nanjing, China, 211166"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5919-7168","authenticated-orcid":true,"given":"Wei-Jun","family":"Kong","sequence":"additional","affiliation":[{"name":"Tongji University , , , ,","place":["Shanghai, China, 200092"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3607-578X","authenticated-orcid":true,"given":"Biao","family":"Yu","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , , ,","place":["Shanghai, China, 200032"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6217-5450","authenticated-orcid":true,"given":"Pan","family":"Fang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , , ,","place":["Shanghai, China, 200032"]},{"name":"Soochow University , , , ,","place":["Suzhou, China, 215123"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,13]]},"reference":[{"key":"2026072808072966800_cit1","doi-asserted-by":"publisher","first-page":"916","DOI":"10.1126\/science.1141448","article-title":"Adapting proteostasis\nfor disease intervention","volume":"319","author":"Balch","year":"2008","journal-title":"Science"},{"key":"2026072808072966800_cit2","doi-asserted-by":"publisher","first-page":"1231","DOI":"10.1083\/jcb.201612111","article-title":"Shaping\nproteostasis at the cellular,\ntissue, and organismal level","volume":"216","author":"Sala","year":"2017","journal-title":"J. Cell Biol."},{"key":"2026072808072966800_cit3","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1013155","article-title":"Proteostasis\nsignatures in human diseases","volume":"21","author":"Lim","year":"2025","journal-title":"PLoS Comput. Biol."},{"key":"2026072808072966800_cit4","doi-asserted-by":"publisher","first-page":"766","DOI":"10.1126\/science.1194160","article-title":"Cell Biology. The\nproteome in balance","volume":"329","author":"Hutt","year":"2010","journal-title":"Science."},{"key":"2026072808072966800_cit5","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1038\/nrm3383","article-title":"Vertebrate\nprotein glycosylation:\ndiversity, synthesis and function","volume":"13","author":"Moremen","year":"2012","journal-title":"Nat. Rev.\nMol. Cell Biol."},{"key":"2026072808072966800_cit6","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1038\/s41580-020-00294-x","article-title":"Global view of human\nprotein glycosylation pathways and functions","volume":"21","author":"Schjoldager","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2026072808072966800_cit7","doi-asserted-by":"publisher","first-page":"100002","DOI":"10.1016\/j.glycos.2025.100002","article-title":"The importance of IgG N-glycosylation\nin Health, Disease,\nand Neonatal hemochromatosis","volume":"1","author":"Thac\u0327i","year":"2025","journal-title":"Glycoscience &\nTherapy."},{"key":"2026072808072966800_cit8","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1038\/s41392-024-01886-1","article-title":"Glycosylation:\nmechanisms, biological\nfunctions and clinical implications","volume":"9","author":"He","year":"2024","journal-title":"Signal\nTransduct Target Ther."},{"key":"2026072808072966800_cit9","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1007\/s10059-010-0159-z","article-title":"Protein\nN-glycosylation, protein\nfolding, and protein quality control","volume":"30","author":"Roth","year":"2010","journal-title":"Mol. 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Commun."},{"key":"2026072808072966800_cit19","doi-asserted-by":"publisher","first-page":"11494","DOI":"10.1002\/anie.202102692","article-title":"Time-Resolved\nand Comprehensive Analysis\nof Surface Glycoproteins Reveals Distinct Responses of Monocytes and\nMacrophages to Bacterial Infection","volume":"60","author":"Suttapitugsakul","year":"2021","journal-title":"Angew. Chem.,\nInt. Ed."},{"key":"2026072808072966800_cit20","doi-asserted-by":"publisher","first-page":"102460","DOI":"10.1016\/j.cbpa.2024.102460","article-title":"Towards chemoenzymatic labeling strategies for profiling\nprotein glycosylation","volume":"80","author":"Tian","year":"2024","journal-title":"Curr. Opin Chem. Biol."},{"key":"2026072808072966800_cit21","doi-asserted-by":"publisher","first-page":"15879","DOI":"10.1021\/jacs.3c02976","article-title":"\u2033Two\nBirds One Stone\u2033 Strategy for the Site-Specific Analysis of\nCore Fucosylation and O-GlcNAcylation","volume":"145","author":"Luo","year":"2023","journal-title":"J. Am.\nChem. Soc."},{"key":"2026072808072966800_cit22","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202513638","article-title":"A Reversible Chemoenzymatic Labeling Strategy for Profiling of Protein\nO-Glucosylation","volume":"64","author":"Zhang","year":"2025","journal-title":"Angew. Chem., Int. 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Engl."},{"key":"2026072808072966800_cit26","doi-asserted-by":"publisher","first-page":"6974","DOI":"10.1002\/anie.200900942","article-title":"Bioorthogonal chemistry: fishing for selectivity in\na sea of functionality","volume":"48","author":"Sletten","year":"2009","journal-title":"Angew. Chem., Int. Ed.\nEngl."},{"key":"2026072808072966800_cit27","doi-asserted-by":"publisher","first-page":"7145","DOI":"10.3390\/molecules18067145","article-title":"Applications of azide-based bioorthogonal click chemistry in glycobiology","volume":"18","author":"Zhang","year":"2013","journal-title":"Molecules."},{"key":"2026072808072966800_cit28","doi-asserted-by":"publisher","first-page":"604","DOI":"10.1007\/s13361-014-1016-7","article-title":"Mass spectrometric\nanalysis of the\ncell surface N-glycoproteome by combining metabolic labeling and click\nchemistry","volume":"26","author":"Smeekens","year":"2015","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"2026072808072966800_cit29","doi-asserted-by":"publisher","first-page":"306","DOI":"10.1039\/D1CB00010A","article-title":"Design and Synthesis\nof Metabolic Chemical Reporters\nfor the Visualization and Identification of Glycoproteins","volume":"2","author":"Pedowitz","year":"2021","journal-title":"RSC Chem. Biol."},{"key":"2026072808072966800_cit30","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1016\/S0076-6879(06)15015-6","article-title":"Metabolic labeling of glycans with azido sugars for\nvisualization and glycoproteomics","volume":"415","author":"Laughlin","year":"2006","journal-title":"Methods Enzymol."},{"key":"2026072808072966800_cit31","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1002\/9780470559277.ch150185","article-title":"Isotope Targeted\nGlycoproteomics (IsoTaG) to Characterize\nIntact, Metabolically Labeled Glycopeptides from Complex Proteomes","volume":"8","author":"Woo","year":"2016","journal-title":"Curr. Protoc Chem. Biol."},{"key":"2026072808072966800_cit32","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202303410","article-title":"Click-iG: Simultaneous\nEnrichment and Profiling of Intact N-linked,\nO-GalNAc, and O-GlcNAcylated Glycopeptides","volume":"62","author":"Liu","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026072808072966800_cit33","doi-asserted-by":"publisher","first-page":"9528","DOI":"10.1039\/C5OB01457C","article-title":"Copper-catalysed azide-alkyne cycloadditions\n(CuAAC): an update","volume":"13","author":"Haldon","year":"2015","journal-title":"Org. Biomol Chem."},{"key":"2026072808072966800_cit34","doi-asserted-by":"publisher","first-page":"2596","DOI":"10.1002\/1521-3773(20020715)41:14&lt;2596::AID-ANIE2596&gt;3.0.CO;2-4","article-title":"A stepwise\nhuisgen cycloaddition process: copper(I)-catalyzed regioselective\n\u2033ligation\u2033 of azides and terminal alkynes","volume":"41","author":"Rostovtsev","year":"2002","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026072808072966800_cit35","doi-asserted-by":"publisher","first-page":"9879","DOI":"10.1002\/anie.200905087","article-title":"Analysis and optimization\nof copper-catalyzed azide-alkyne cycloaddition for bioconjugation","volume":"48","author":"Hong","year":"2009","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026072808072966800_cit36","doi-asserted-by":"publisher","first-page":"15046","DOI":"10.1021\/ja044996f","article-title":"A strain-promoted [3 + 2] azide-alkyne\ncycloaddition for covalent modification of biomolecules in living\nsystems","volume":"126","author":"Agard","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026072808072966800_cit37","doi-asserted-by":"publisher","first-page":"1912","DOI":"10.1002\/cbic.201100117","article-title":"Strain-promoted\nalkyne-azide cycloadditions (SPAAC) reveal new features of glycoconjugate\nbiosynthesis","volume":"12","author":"Mbua","year":"2011","journal-title":"ChemBioChem"},{"key":"2026072808072966800_cit38","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1007\/s41061-016-0016-4","article-title":"Strain-Promoted 1,3-Dipolar Cycloaddition\nof Cycloalkynes and Organic Azides","volume":"374","author":"Dommerholt","year":"2016","journal-title":"Top Curr.\nChem. 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Aspects Med."},{"key":"2026072808072966800_cit44","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1007\/s00262-025-03979-4","article-title":"Prostaglandin F2 receptor negative\nregulator as a potential\ntarget for chimeric antigen receptor-T cell therapy for glioblastoma","volume":"74","author":"Kuroda","year":"2025","journal-title":"Cancer Immunol Immunother."},{"key":"2026072808072966800_cit45","doi-asserted-by":"publisher","first-page":"13612","DOI":"10.1073\/pnas.0606424103","article-title":"Zip14 (Slc39a14)\nmediates non-transferrin-bound iron uptake into cells","volume":"103","author":"Liuzzi","year":"2006","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026072808072966800_cit46","doi-asserted-by":"publisher","first-page":"969","DOI":"10.2147\/OTT.S40241","article-title":"The potential\nroles of hepatocyte growth factor (HGF)-MET pathway inhibitors in\ncancer treatment","volume":"7","author":"Appleman","year":"2014","journal-title":"Onco Targets Ther."}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.6c02598","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/29\/21697\/65885019\/acs.analchem.6c02598.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/29\/21697\/65885019\/acs.analchem.6c02598.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T12:22:01Z","timestamp":1785241321000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/98\/29\/21697\/5207051\/Chemically-Cleavable-Enrichment-Enables-Site"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,13]]},"references-count":46,"journal-issue":{"issue":"29","published-online":{"date-parts":[[2026,7,13]]},"published-print":{"date-parts":[[2026,7,28]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.6c02598","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,13]]}}}