title: PUGNAc treatment provokes globotetraosylceramide accumulation in human umbilical vein endothelial cells.
abstract: PUGNAc is a well-investigated inhibitor for protein-O-GlcNAcase, whereas recent investigations showed that PUGNAc had a broad range as inhibitor for cellular β-hexosaminidases. Here we report that PUGNAc treatment provokes globotetraosylceramide (Gb4Cer) accumulation in human umbilical vein endothelial cells (HUVEC). HPLC analysis and a quantitative ELISA using newly developed anti-Gb4Cer monoclonal antibody revealed that PUGNAc treatment specifically increased the expression of Gb4Cer among glycosphingolipids expressed in HUVEC. Although the effect was weaker than PUGNAc, an O-GlcNAcase selective inhibitor (Thiamet-G) treatment also increased Gb4Cer levels in HUVEC. Furthermore, both of PUGNAc and Thiamet-G treatment up-regulated the expression levels of α-1,4-galactosyltransferase/Gb3Cer synthase gene which encodes a key enzyme in Gb4Cer synthesis. These results indicate that protein-O-GlcNAcylation can regulate the expression levels of cellular Gb4Cer.
authors.affiliation: Bio-Design Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. Electronic address: t-okuda@aist.go.jp.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Okuda
mesh_terms: D000117:Acetylglucosamine / Q000008:administration & dosage / Q000031:analogs & derivatives*; D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D005915:Globosides / Q000096:biosynthesis*; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D010091:Oximes / Q000008:administration & dosage*; D048448:Phenylcarbamates / Q000008:administration & dosage*; D014471:Umbilical Veins / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D005915:Globosides; D010091:Oximes; D048448:Phenylcarbamates; C024032:globotetraosylceramide; C068836:N-acetylglucosaminono-1,5-lactone O-(phenylcarbamoyl)oxime; D000117:Acetylglucosamine
keywords: Globoside; Glycosphingolipid; O-GlcNAc; Sandhoff disease; Vascular endothelial cell; β-hexosaminidase