{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T03:43:04Z","timestamp":1767843784510,"version":"3.49.0"},"reference-count":26,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2023,4,11]],"date-time":"2023-04-11T00:00:00Z","timestamp":1681171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Japan Agency for Medical Research and Development","award":["21fk0210079h0002"],"award-info":[{"award-number":["21fk0210079h0002"]}]},{"name":"Japan Agency for Medical Research and Development","award":["22H02967"],"award-info":[{"award-number":["22H02967"]}]},{"name":"Ministry of Education, Culture, Sports, Science and Technology of Japan (JSPS KAKENHI)","award":["21fk0210079h0002"],"award-info":[{"award-number":["21fk0210079h0002"]}]},{"name":"Ministry of Education, Culture, Sports, Science and Technology of Japan (JSPS KAKENHI)","award":["22H02967"],"award-info":[{"award-number":["22H02967"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Fucosylated proteins are widely used as biomarkers of cancer and inflammation. Fucosylated alpha-fetoprotein (AFP-L3) is a specific biomarker for hepatocellular carcinoma. We previously showed that increases in serum AFP-L3 levels depend on increased expression of fucosylation-regulatory genes and abnormal transport of fucosylated proteins in cancer cells. In normal hepatocytes, fucosylated proteins are selectively secreted in the bile duct but not blood. In cases of cancer cells without cellular polarity, this selective secretion system is destroyed. Here, we aimed to identify cargo proteins involved in the selective secretion of fucosylated proteins, such as AFP-L3, into bile duct-like structures in HepG2 hepatoma cells, which have cellular polarity like, in part, normal hepatocytes. \u03b11-6 Fucosyltransferase (FUT8) is a key enzyme to synthesize core fucose and produce AFP-L3. Firstly, we knocked out the FUT8 gene in HepG2 cells and investigated the effects on the secretion of AFP-L3. AFP-L3 accumulated in bile duct-like structures in HepG2 cells, and this phenomenon was diminished by FUT8 knockout, suggesting that HepG2 cells have cargo proteins for AFP-L3. To identify cargo proteins involved in the secretion of fucosylated proteins in HepG2 cells, immunoprecipitation and the proteomic Strep-tag system experiments followed by mass spectrometry analyses were performed. As a result of proteomic analysis, seven kinds of lectin-like molecules were identified, and we selected vesicular integral membrane protein gene VIP36 as a candidate of the cargo protein that interacts with the \u03b11-6 fucosylation (core fucose) on N-glycan according to bibliographical consideration. Expectedly, the knockout of the VIP36 gene in HepG2 cells suppressed the secretion of AFP-L3 and other fucosylated proteins, such as fucosylated alpha-1 antitrypsin, into bile duct-like structures. We propose that VIP36 could be a cargo protein involved in the apical secretion of fucosylated proteins in HepG2 cells.<\/jats:p>","DOI":"10.3390\/ijms24087037","type":"journal-article","created":{"date-parts":[[2023,4,11]],"date-time":"2023-04-11T02:02:33Z","timestamp":1681178553000},"page":"7037","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Vesicular Integral-Membrane Protein 36 Is Involved in the Selective Secretion of Fucosylated Proteins into Bile Duct-like Structures in HepG2 Cells"],"prefix":"10.3390","volume":"24","author":[{"given":"Mizuki","family":"Muranaka","sequence":"first","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1996-7740","authenticated-orcid":false,"given":"Shinji","family":"Takamatsu","sequence":"additional","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tsunenori","family":"Ouchida","sequence":"additional","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuri","family":"Kanazawa","sequence":"additional","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jumpei","family":"Kondo","sequence":"additional","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tsutomu","family":"Nakagawa","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, School of Pharmaceutical Sciences, Health Sciences University of Hokkaido, 1757 Kanazawa, Toubetsu-cho, Ishikari-gun 061-0293, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuriko","family":"Egashira","sequence":"additional","affiliation":[{"name":"Bio-Diagnostic Reagent Technology Center, Sysmex Corporation, 4-3-2 Takatsukadai, Nishi-ku, Kobe 651-2271, Hyogo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koji","family":"Fukagawa","sequence":"additional","affiliation":[{"name":"Bio-Diagnostic Reagent Technology Center, Sysmex Corporation, 4-3-2 Takatsukadai, Nishi-ku, Kobe 651-2271, Hyogo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0241-8788","authenticated-orcid":false,"given":"Jianguo","family":"Gu","sequence":"additional","affiliation":[{"name":"Division of Regulatory Glycobiology, Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, 4-4-1 Komatsushima, Aobaku, Sendai 981-8558, Miyagi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toru","family":"Okamoto","sequence":"additional","affiliation":[{"name":"Institute for Advanced Co-Creation Studies, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshihiro","family":"Kamada","sequence":"additional","affiliation":[{"name":"Department of Advanced Metabolic Hepatology, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9430-2362","authenticated-orcid":false,"given":"Eiji","family":"Miyoshi","sequence":"additional","affiliation":[{"name":"Department of Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, 1-7 Yamada-oka, Suita 565-0871, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1146\/annurev.bi.57.070188.004033","article-title":"Glycobiology","volume":"57","author":"Rademacher","year":"1988","journal-title":"Annu. 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Protoc."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/8\/7037\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:13:35Z","timestamp":1760123615000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/8\/7037"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,11]]},"references-count":26,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2023,4]]}},"alternative-id":["ijms24087037"],"URL":"https:\/\/doi.org\/10.3390\/ijms24087037","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,11]]}}}