{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T03:13:54Z","timestamp":1793157234218,"version":"4.5.0"},"reference-count":0,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2025,3,10]],"date-time":"2025-03-10T00:00:00Z","timestamp":1741564800000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Mucin type O-glycan core elongation is typically performed by the C1GALT1, B3GNT6, and ST6GalNAc-I\/-II O-glycosyltransferases. These enzymes target the Tn antigen (GalNAc-O-Thr\/Ser) dictating the fate of O-glycan elongation, playing important roles in health and disease. Changes in transferase expression and glycan structure are commonly associated with diseases such as cancer, Tn-syndrome, and ulcerative colitis. Despite their significance, their substrate specificities and their biological roles remain elusive. Here, we examine the roles of flanking glycopeptide substrate charge using a library of differently charged glycopeptides and a small library of PSGL-1 Thr57 based charged glycopeptides. We found that C1GALT1 was most influenced by flanking charge preferring negatively charged substrates, while B3GNT6 and ST6GalNAc-II were less influenced, showing unique N- and C-terminal charge preferences. Interestingly, ST6GalNAc-I was not influenced by flanking charge. These charge specificities were further maintained against the charged PSGL-1 glycopeptides, although ST6GalNAc-I showed an increased preference towards a remote N-terminal positive charge. The observed charge preferences were to a large part driven by substrate interactions with the electrostatic surface of the transferase. We propose that negative flanking charge may assist C1GALT1 in targeting key glycosites such as in PSGL-1 and podoplanin. Our findings are consistent with a Golgi hierarchy, where the cis-Golgi localized GalNAc-Ts and C1GALT1 determine the site and thus fate of glycosylation, while the trans-Golgi less-specific ST6GalNAc-I provides a final capping function. This characterization of substrate charge preference furthers our understanding of how these enzymes select their substrates and may contribute to our understanding of their biological roles.<\/jats:p>","DOI":"10.1093\/glycob\/cwaf014","type":"journal-article","created":{"date-parts":[[2025,3,10]],"date-time":"2025-03-10T19:45:17Z","timestamp":1741635917000},"source":"Crossref","is-referenced-by-count":3,"title":["Charge matters: how flanking substrate charge modulates O-glycan Core elongation"],"prefix":"10.1093","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2869-6970","authenticated-orcid":false,"given":"Collin J","family":"Ballard","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7111-8331","authenticated-orcid":false,"given":"Matthew R","family":"Smutny","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1215-087X","authenticated-orcid":false,"given":"Lam D","family":"Chau","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1818-9806","authenticated-orcid":false,"given":"Collin K","family":"Wong","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]},{"name":"Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic , Cleveland, OH 44195 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8038-3438","authenticated-orcid":false,"given":"Haley M","family":"Aharoni","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]},{"name":"Department of Pharmacology, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0465-659X","authenticated-orcid":false,"given":"Hana K","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0649-1368","authenticated-orcid":false,"given":"Digantkumar G","family":"Chapla","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia , Athens, Georgia 30602 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramon","family":"Hurtado-Guerrero","sequence":"additional","affiliation":[{"name":"The Institute for Biocomputation and Physics of Complex Systems (BIFI), University of Zaragoza , Mariano Esquillor s\/n, Campus Rio Ebro, Edificio I+D, Zaragoza 50018 ,","place":["Spain"]},{"name":"Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen , Blegdamsvej 3, DK-2200, Copenhagen N ,","place":["Denmark"]},{"name":"Fundaci\u00f3n ARAID , Zaragoza 50018 ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1768-582X","authenticated-orcid":false,"given":"Kelley W","family":"Moremen","sequence":"additional","affiliation":[{"name":"Complex Carbohydrate Research Center, University of Georgia , Athens, Georgia 30602 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7919-7613","authenticated-orcid":false,"given":"Thomas A","family":"Gerken","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Case Western Reserve University , Cleveland, OH 44106 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,3,10]]},"container-title":["Glycobiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/glycob\/advance-article-pdf\/doi\/10.1093\/glycob\/cwaf014\/62369688\/cwaf014.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/glycob\/advance-article-pdf\/doi\/10.1093\/glycob\/cwaf014\/62369688\/cwaf014.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,10]],"date-time":"2025-03-10T19:45:17Z","timestamp":1741635917000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/glycob\/advance-article\/doi\/10.1093\/glycob\/cwaf014\/8068942"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,10]]},"references-count":0,"URL":"https:\/\/doi.org\/10.1093\/glycob\/cwaf014","relation":{},"ISSN":["1460-2423"],"issn-type":[{"value":"1460-2423","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,10]]},"article-number":"cwaf014"}}