{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T05:12:40Z","timestamp":1762060360692,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2022,8,2]],"date-time":"2022-08-02T00:00:00Z","timestamp":1659398400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Grant-in-Aid for Scientific Research (C), JSPS","award":["JP18K06649","JP17H06417","JP20gm0910005j0006"],"award-info":[{"award-number":["JP18K06649","JP17H06417","JP20gm0910005j0006"]}]},{"name":"MEXT KAKENHI","award":["JP18K06649","JP17H06417","JP20gm0910005j0006"],"award-info":[{"award-number":["JP18K06649","JP17H06417","JP20gm0910005j0006"]}]},{"DOI":"10.13039\/100009619","name":"AMED-CREST","doi-asserted-by":"publisher","award":["JP18K06649","JP17H06417","JP20gm0910005j0006"],"award-info":[{"award-number":["JP18K06649","JP17H06417","JP20gm0910005j0006"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Ceramide transport protein (CERT) mediates ceramide transfer from the endoplasmic reticulum to the Golgi for sphingomyelin (SM) biosynthesis. CERT is inactivated by multiple phosphorylation at the serine-repeat motif (SRM), and mutations that impair the SRM phosphorylation are associated with a group of inherited intellectual disorders in humans. It has been suggested that the N-terminal phosphatidylinositol 4-monophosphate [PtdIns(4)P] binding domain and the C-terminal ceramide-transfer domain of CERT physically interfere with each other in the SRM phosphorylated state, thereby repressing the function of CERT; however, it remains unclear which regions in CERT are involved in the SRM phosphorylation-dependent repression of CERT. Here, we identified a previously uncharacterized cluster of lysine\/arginine residues that were predicted to be located on the outer surface of a probable coiled-coil fold in CERT. Substitutions of the basic amino acids in the cluster with alanine released the SRM-dependent repression of CERT activities, i.e., the synthesis of SM, PtdIns(4)P-binding, vesicle-associated membrane protein-associated protein (VAP) binding, ceramide-transfer activity, and localization to the Golgi, although the effect on SM synthesis activity was only partially compromised by the alanine substitutions, which moderately destabilized the trimeric status of CERT. These results suggest that the basic amino acid cluster in the coiled-coil region is involved in the regulation of CERT function.<\/jats:p>","DOI":"10.3390\/ijms23158576","type":"journal-article","created":{"date-parts":[[2022,8,2]],"date-time":"2022-08-02T21:41:37Z","timestamp":1659476497000},"page":"8576","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Involvement of a Cluster of Basic Amino Acids in Phosphorylation-Dependent Functional Repression of the Ceramide Transport Protein CERT"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0002-2619","authenticated-orcid":false,"given":"Asako","family":"Goto","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2810-5296","authenticated-orcid":false,"given":"Daichi","family":"Egawa","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1266-5400","authenticated-orcid":false,"given":"Nario","family":"Tomishige","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshiyuki","family":"Yamaji","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kentaro","family":"Shimasaki","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keigo","family":"Kumagai","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1383-2781","authenticated-orcid":false,"given":"Kentaro","family":"Hanada","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"},{"name":"Department of Quality Assurance, Radiation Safety, and Information System, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2515256420959514","DOI":"10.1177\/2515256420959514","article-title":"Sphingolipid Metabolism at the ER-Golgi Contact Zone and Its Impact on Membrane Trafficking","volume":"3","author":"Goto","year":"2020","journal-title":"Contact"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1194\/jlr.R085324","article-title":"Lipid Transfer Proteins Rectify Inter-Organelle Flux and Accurately Deliver Lipids at Membrane Contact Sites","volume":"59","author":"Hanada","year":"2018","journal-title":"J. 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