title: Interaction of FAM5C with UDP-glucose:glycoprotein glucosyltransferase 1 (UGGT1): Implication of N-glycosylation in FAM5C secretion.
abstract: N-glycosylation of proteins is important for protein folding and function. We have recently reported that FAM5C/BRINP3 contributes to the tumor necrosis factor-α-induced expression of leukocyte adhesion molecules in vascular endothelial cells (ECs). However, regulatory mechanism of the FAM5C biosynthesis is poorly understood. Co-immunoprecipitation assay revealed the interaction of FAM5C with UDP-glucose:glycoprotein glucosyltransferase 1 (UGGT1), a glycoprotein folding-sensor enzyme. FAM5C ectopically expressed in HEK293 cells was localized to the endoplasmic reticulum and co-localized with endogenously expressed UGGT1. Molecular size of FAM5C was reduced by treatment with N-glycosidase F and in FAM5C-expressing cells cultured in the presence of the N-glycosylation inhibitor tunicamycin. FAM5C was secreted by the cells and the secretion of FAM5C was blocked by tunicamycin. Among six potential N-glycosylation sites, the potential site at Asn168was not N-glycosylated, and Asn337, Asn456, Asn562, Asn609, and Asn641mutants were poorly secreted by the cells. These results demonstrated that FAM5C is an N-glycosylated protein and N-glycosylation is necessary for the secretion of FAM5C.
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan; Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Terao
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan.
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Fujita
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan.
authors.forename: Sayo
authors.initials: S
authors.lastname: Horibe
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan; Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Sato
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Minami
authors.affiliation: Laboratory of Physiological Chemistry, College of Pharmaceutical Sciences, Matsuyama University, Matsuyama, Ehime 790-8578, Japan.
authors.forename: Miwako
authors.initials: M
authors.lastname: Kobayashi
authors.affiliation: Laboratory of Physiological Chemistry, College of Pharmaceutical Sciences, Matsuyama University, Matsuyama, Ehime 790-8578, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Matsuoka
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan; Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Sasaki
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan.
authors.forename: Seimi
authors.initials: S
authors.lastname: Satomi-Kobayashi
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan.
authors.forename: Ken-Ichi
authors.initials: KI
authors.lastname: Hirata
authors.affiliation: Department of Medical Pharmaceutics, Kobe Pharmaceutical University, Kobe, Hyogo 658-8558, Japan; Division of Signal Transduction, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, Kobe, Hyogo 650-0017, Japan. Electronic address: rikitake@kobepharma-u.ac.jp.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Rikitake
mesh_terms: D001216:Asparagine / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D014415:Tunicamycin / Q000494:pharmacology; D000099237:UDP-Galactose Translocators
chemical_list: D001216:Asparagine; D004268:DNA-Binding Proteins; D005964:Glucosyltransferases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D014415:Tunicamycin; C528897:BRINP3 protein, human; C000731541:UGGT1 protein, human; D000099237:UDP-Galactose Translocators
keywords: BRINP3; Endothelial cells; FAM5C; N-glycosylation; UGGT1