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Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase.
Gludovacz, Elisabeth; Maresch, Daniel; Lopes de Carvalho, Leonor; Puxbaum, Verena; Baier, Laurenz J; Sützl, Leander; Guédez, Gabriela; Grünwald-Gruber, Clemens; Ulm, Barbara; Pils, Sophie; Ristl, Robin; Altmann, Friedrich; Jilma, Bernd; Salminen, Tiina A; Borth, Nicole; Boehm, Thomas.
Afiliação
  • Gludovacz E; From the Departments of Biotechnology.
  • Maresch D; the Departments of Clinical Pharmacology and.
  • Lopes de Carvalho L; Chemistry, and.
  • Puxbaum V; the Structural Bioinformatics Laboratory, Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, Tykistökatu 6A, 20520 Turku, Finland.
  • Baier LJ; From the Departments of Biotechnology.
  • Sützl L; From the Departments of Biotechnology.
  • Guédez G; Food Science and Technology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
  • Grünwald-Gruber C; the Structural Bioinformatics Laboratory, Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, Tykistökatu 6A, 20520 Turku, Finland.
  • Ulm B; Chemistry, and.
  • Pils S; Obstetrics and Gynecology, and.
  • Ristl R; Obstetrics and Gynecology, and.
  • Altmann F; the Section for Medical Statistics (IMS), Center of Medical Statistics, Informatics and Intelligent Systems, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria, and.
  • Jilma B; Chemistry, and.
  • Salminen TA; the Departments of Clinical Pharmacology and.
  • Borth N; the Structural Bioinformatics Laboratory, Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, Tykistökatu 6A, 20520 Turku, Finland.
  • Boehm T; From the Departments of Biotechnology, nicole.borth@boku.ac.at.
J Biol Chem ; 293(3): 1070-1087, 2018 01 19.
Article em En | MEDLINE | ID: mdl-29187599
N-Glycosylation plays a fundamental role in many biological processes. Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. Mutations of Asn-168, Asn-538, and Asn-745 reduced rhDAO secretion by 13, 71, and 32%, respectively. Asn-538/745 double and Asn-168/538/745 triple substitutions reduced rhDAO secretion by 85 and 94%. Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oligossacarídeos / Polissacarídeos / Amina Oxidase (contendo Cobre) Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oligossacarídeos / Polissacarídeos / Amina Oxidase (contendo Cobre) Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article