title: Expression of long-form N-acetylglucosamine-6-O-sulfotransferase 1 in human high endothelial venules.
abstract: Two members of the N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) family, GlcNAc6ST-1 and GlcNAc6ST-2, function in the biosynthesis of 6-sulfo sialyl Lewis X-capped glycoproteins expressed on high endothelial venules (HEVs) in secondary lymphoid organs. Thus, both enzymes play a critical role in L-selectin-expressing lymphocyte homing. Human GlcNAc6ST-1 is encoded by a 1593-bp open reading frame exhibiting two 5' in-frame methionine codons spaced 141 bp apart. Both resemble the consensus sequence for translation initiation. Thus, it has been hypothesized that both long and short forms of GlcNAc6ST-1 may be present, although endogenous expression of either form has not been confirmed in humans. Here, the authors developed an antibody recognizing amino acid residues between the first two human GlcNAc6ST-1 methionines. This antibody specifically recognizes the long form of the enzyme, a finding validated by Western blot analysis and immunofluorescence cytochemistry of HeLa cells misexpressing long and/or short forms of human GlcNAc6ST-1. Using this antibody, the authors carried out immunofluorescence histochemistry of human lymph node tissue sections and found endogenous expression of the long form of the enzyme in human tissue, predominantly in the trans-Golgi network of endothelial cells that form HEVs.
authors.affiliation: Department of Molecular Pathology, Shinshu University Graduate School of Medicine, Matsumoto, Japan.
authors.forename: Maiko
authors.initials: M
authors.lastname: Fujiwara
authors.forename: Motohiro
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Hitomi
authors.initials: H
authors.lastname: Hoshino
authors.forename: Kenji
authors.initials: K
authors.lastname: Uchimura
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakada
authors.forename: Junya
authors.initials: J
authors.lastname: Masumoto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Sakai
authors.forename: Minoru
authors.initials: M
authors.lastname: Fukuda
authors.forename: Jun
authors.initials: J
authors.lastname: Nakayama
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies; D001483:Base Sequence; D015153:Blotting, Western; D004730:Endothelium, Vascular / Q000201:enzymology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D008198:Lymph Nodes / Q000098:blood supply; D008969:Molecular Sequence Data; D015238:Sulfotransferases / Q000276:immunology / Q000378:metabolism*; D014699:Venules / Q000201:enzymology* / Q000648:ultrastructure; D000097811:Carbohydrate Sulfotransferases
chemical_list: D000906:Antibodies; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases