title: Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells.
abstract: DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. By combining these methods, a high heterogeneity of DENV N-glycans was found. Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells.
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: Y
authors.initials: Y
authors.lastname: Lei
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, 229 Taibai Beilu, Xi'an 710069, China.
authors.forename: H
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: Y
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: W
authors.initials: W
authors.lastname: Ye
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, 229 Taibai Beilu, Xi'an 710069, China.
authors.forename: W
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Infectious Diseases and Center of liver Diseases, Tangdu Hospital, the Fourth Military Medical University, 569 Xinsi Road, Xi'an 710038, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Jia
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: Z
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, 229 Taibai Beilu, Xi'an 710069, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Microbiology, School of Preclinical Medicine, The Fourth Military Medical University, 169 Changle Xi Road, Xian, Shaanxi 710032 China.
authors.forename: F
authors.initials: F
authors.lastname: Zhang
mesh_terms: D000330:Aedes / Q000166:cytology / Q000821:virology; D000818:Animals; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000821:virology; D003716:Dengue Virus / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005643:Fucose / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006860:Hydrogen Bonding; D037181:Lectins, C-Type / Q000737:chemistry / Q000378:metabolism; D008358:Mannose / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D014771:Virion / Q000737:chemistry / Q000378:metabolism*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D015815:Cell Adhesion Molecules; D005643:Fucose; D037181:Lectins, C-Type; D008358:Mannose; D009844:Oligosaccharides; D011134:Polysaccharides; D011956:Receptors, Cell Surface; D014759:Viral Envelope Proteins; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin