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Trastuzumab\u2019s target epitope is localized within the extracellular domain of the oncogenic cell surface receptor tyrosine kinase (RTK) ErbB2, which is known to undergo extensive <jats:italic>N<\/jats:italic>-linked glycosylation. However, the site-specific glycan repertoire of ErbB2, as well as the detailed molecular mechanisms through which specific aberrant glycan signatures functionally impact the malignant features of ErbB2-addicted GC cells, including the acquisition of trastuzumab resistance, remain elusive. Here, we demonstrate that ErbB2 is modified with both \u03b12,6- and \u03b12,3-sialylated glycan structures in GC clinical specimens. In-depth mass spectrometry-based glycomic and glycoproteomic analysis of ErbB2\u2019s ectodomain disclosed a site-specific glycosylation profile in GC cells, in which the ST6Gal1 sialyltransferase specifically targets ErbB2 <jats:italic>N<\/jats:italic>-glycosylation sites occurring within the receptor\u2019s trastuzumab-binding domain. Abrogation of ST6Gal1 expression reshaped the cellular and ErbB2-specific glycomes, expanded the cellular half-life of the ErbB2 receptor, and sensitized ErbB2-dependent GC cells to trastuzumab-induced cytotoxicity through the stabilization of ErbB dimers at the cell membrane, and the decreased activation of both ErbB2 and EGFR RTKs. Overall, our data demonstrates that ST6Gal1-mediated aberrant \u03b12,6-sialylation actively tunes the resistance of ErbB2-driven GC cells to trastuzumab.<\/jats:p>","DOI":"10.1038\/s41388-021-01801-w","type":"journal-article","created":{"date-parts":[[2021,5,4]],"date-time":"2021-05-04T12:04:09Z","timestamp":1620129849000},"page":"3719-3733","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":58,"title":["ST6Gal1 targets the ectodomain of ErbB2 in a site-specific manner and regulates gastric cancer cell sensitivity to trastuzumab"],"prefix":"10.1038","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8641-8368","authenticated-orcid":false,"given":"Henrique O.","family":"Duarte","sequence":"first","affiliation":[]},{"given":"Joana G.","family":"Rodrigues","sequence":"additional","affiliation":[]},{"given":"Catarina","family":"Gomes","sequence":"additional","affiliation":[]},{"given":"Paul J.","family":"Hensbergen","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1689-0442","authenticated-orcid":false,"given":"Agnes L. 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