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Site-specific N-glycosylation analysis of animal cell culture-derived Zika virus proteins.
Pralow, Alexander; Nikolay, Alexander; Leon, Arnaud; Genzel, Yvonne; Rapp, Erdmann; Reichl, Udo.
Afiliação
  • Pralow A; Bioprocess Engineering Group, Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany.
  • Nikolay A; Bioprocess Engineering Group, Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany.
  • Leon A; Valneva SE, Saint-Herblain, France.
  • Genzel Y; Bioprocess Engineering Group, Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany.
  • Rapp E; Bioprocess Engineering Group, Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany. rapp@mpi-magdeburg.mpg.de.
  • Reichl U; glyXera GmbH, Magdeburg, Germany. rapp@mpi-magdeburg.mpg.de.
Sci Rep ; 11(1): 5147, 2021 03 04.
Article em En | MEDLINE | ID: mdl-33664361
Here, we present for the first time, a site-specific N-glycosylation analysis of proteins from a Brazilian Zika virus (ZIKV) strain. The virus was propagated with high yield in an embryo-derived stem cell line (EB66, Valneva SE), and concentrated by g-force step-gradient centrifugation. Subsequently, the sample was proteolytically digested with different enzymes, measured via a LC-MS/MS-based workflow, and analyzed in a semi-automated way using the in-house developed glyXtoolMS software. The viral non-structural protein 1 (NS1) was glycosylated exclusively with high-mannose structures on both potential N-glycosylation sites. In case of the viral envelope (E) protein, no specific N-glycans could be identified with this method. Nevertheless, N-glycosylation could be proved by enzymatic de-N-glycosylation with PNGase F, resulting in a strong MS-signal of the former glycopeptide with deamidated asparagine at the potential N-glycosylation site N444. This confirmed that this site of the ZIKV E protein is highly N-glycosylated but with very high micro-heterogeneity. Our study clearly demonstrates the progress made towards site-specific N-glycosylation analysis of viral proteins, i.e. for Brazilian ZIKV. It allows to better characterize viral isolates, and to monitor glycosylation of major antigens. The method established can be applied for detailed studies regarding the impact of protein glycosylation on antigenicity and human pathogenicity of many viruses including influenza virus, HIV and corona virus.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Glicosilação / Proteínas não Estruturais Virais / Zika virus / Infecção por Zika virus Limite: Animals / Humans País/Região como assunto: America do sul / Brasil Idioma: En Revista: Sci Rep Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Alemanha País de publicação: Reino Unido
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Glicosilação / Proteínas não Estruturais Virais / Zika virus / Infecção por Zika virus Limite: Animals / Humans País/Região como assunto: America do sul / Brasil Idioma: En Revista: Sci Rep Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Alemanha País de publicação: Reino Unido