N-glycosylation profiling of porcine reproductive and respiratory syndrome virus envelope glycoprotein 5

Juan Li, Shujuan Tao, Ron Orlando, Michael P. Murtaugh

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is a positive-sense ssRNA virus whose envelope contains four glycoproteins and three nonglycosylated proteins. Glycans of major envelope glycoprotein 5 (GP5) are proposed as important for virus assembly and entry into permissive cells. Structural characterization of GP5 glycans would facilitate the mechanistic understanding of these processes. Thus, we purified the PRRSV type 2 prototype strain, VR2332, and analyzed the virion-associated glycans by both biochemical and mass spectrometric methods. Endoglycosidase digestion showed that GP5 was the primary protein substrate, and that the carbohydrate moieties were primarily complex-type N-glycans. Mass spectrometric analysis (HPLC-ESI-MS/MS) of GP5 N-glycans revealed an abundance of N-acetylglucosamine (GlcNAc) and N-acetyllactosamine (LacNAc) oligomers in addition to sialic acids. GlcNAc and LacNAc accessibility to ligands was confirmed by lectin co-precipitation. Our findings help to explain PRRSV infection of cells lacking sialoadhesin and provide a glycan database to facilitate molecular structural studies of PRRSV.

Original languageEnglish (US)
Pages (from-to)86-98
Number of pages13
JournalVirology
Volume478
DOIs
StatePublished - Apr 1 2015

Bibliographical note

Funding Information:
The work was funded in part by Grant number 09-227 from the U.S. National Pork Board , the National Institutes of Health/NCRR-funded Integrated Technology Resource for Biomedical Glycomics ( P41 RR018502 ), and USDA NIFA multistate project MIN 63-084 .

Publisher Copyright:
© 2015.

Keywords

  • GP5
  • GlcNAc
  • LacNAc
  • Lectin
  • Mass spectrometry
  • N-glycan
  • PRRSV
  • Veterinary virology
  • Virion structure

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