{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T10:04:53Z","timestamp":1787997893419,"version":"build-2784847793"},"reference-count":39,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2024,1,22]],"date-time":"2024-01-22T00:00:00Z","timestamp":1705881600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/100009633","name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","doi-asserted-by":"publisher","award":["R01HD104808"],"award-info":[{"award-number":["R01HD104808"]}],"id":[{"id":"10.13039\/100009633","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100017749","name":"Cardiovascular center Medical College of Wisconsin","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100017749","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,4,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>O-GlcNAcylation is a dynamic modulator of signaling pathways, equal in magnitude to the widely studied phosphorylation. With the rapid development of tools for its detection at the single protein level, the O-GlcNAc modification rapidly emerged as a novel diagnostic and therapeutic target in human diseases. Yet, mapping the human O-GlcNAcome in various tissues is essential for generating relevant biomarkers. In this study, we used human banked tissue as a sample source to identify O-GlcNAcylated protein targets relevant to human diseases. Using human term placentas, we propose (1) a method to clean frozen banked tissue of blood proteins; (2) an optimized protocol for the enrichment of O-GlcNAcylated proteins using immunoaffinity purification; and (3) a bioinformatic workflow to identify the most promising O-GlcNAc targets. As a proof-of-concept, we used 45\u00a0mg of banked placental samples from two pregnancies to generate intracellular protein extracts depleted of blood protein. Then, antibody-based O-GlcNAc enrichment on denatured samples yielded over 2000 unique HexNAc PSMs and 900 unique sites using 300\u00a0\u03bcg of protein lysate. Due to efficient sample cleanup, we also captured 82 HexNAc proteins with high placental expression. Finally, we provide a bioinformatic tool (CytOVS) to sort the HexNAc proteins based on their cellular localization and extract the most promising O-GlcNAc targets to explore further. To conclude, we provide a simple 3-step workflow to generate a manageable list of O-GlcNAc proteins from human tissue and improve our understanding of O-GlcNAcylation\u2019s role in health and diseases.<\/jats:p>","DOI":"10.1093\/glycob\/cwae005","type":"journal-article","created":{"date-parts":[[2024,1,22]],"date-time":"2024-01-22T23:20:35Z","timestamp":1705965635000},"source":"Crossref","is-referenced-by-count":10,"title":["Studying the <i>O<\/i>-GlcNAcome of human placentas using banked tissue samples"],"prefix":"10.1093","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8917-6196","authenticated-orcid":false,"given":"Sarai","family":"Luna","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Medical College of Wisconsin , 8701 Watertown Plank Rd., Milwaukee, WI 53226 , United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4908-1588","authenticated-orcid":false,"given":"Florian","family":"Malard","sequence":"additional","affiliation":[{"name":"INSERM U1212 , CNRS UMR5320, ARNA Laboratory, , 146 rue L\u00e9o Saignat, 33000 Bordeaux , France"},{"name":"University of Bordeaux , CNRS UMR5320, ARNA Laboratory, , 146 rue L\u00e9o Saignat, 33000 Bordeaux , France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7287-5450","authenticated-orcid":false,"given":"Michaela","family":"Pereckas","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical College of Wisconsin , 8701 Watertown Plank Rd., Milwaukee, WI 53226 , United 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