Merck
  • Home
  • Search Results
  • N-glycosylation profiling of porcine reproductive and respiratory syndrome virus envelope glycoprotein 5.

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

Virology (2015-03-03)
Juan Li, Shujuan Tao, Ron Orlando, Michael P Murtaugh
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acrylamide, for molecular biology, ≥99% (HPLC)
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
Sigma-Aldrich
Sodium bicarbonate, Hybri-Max, powder, suitable for hybridoma, ≥99.5%
Sigma-Aldrich
Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Acrylamide, purum, ≥98.0% (GC)
Sigma-Aldrich
Acetic acid, for luminescence, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Citrate Concentrated Solution, BioUltra, for molecular biology, 1 M in H2O
Sigma-Aldrich
DL-Dithiothreitol solution, BioUltra, for molecular biology, ~1 M in H2O
Supelco
Acetic acid, analytical standard
Sigma-Aldrich
Acrylamide, for Northern and Southern blotting, powder blend
Sigma-Aldrich
Sodium bicarbonate, tested according to Ph. Eur.
Sigma-Aldrich
Citrate Concentrated Solution, BioReagent, suitable for coagulation assays, 4 % (w/v)
Sigma-Aldrich
Acetic acid, natural, ≥99.5%, FG
Sigma-Aldrich
Acetic acid, ≥99.5%, FCC, FG
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
Sigma-Aldrich
Tris(tert-butoxy)silanol, packaged for use in deposition systems
Sigma-Aldrich
HEPES buffer solution, 1 M in H2O
Sigma-Aldrich
Tris(tert-butoxy)silanol, 99.999%
Sigma-Aldrich
Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid
Supelco
Acrylamide, analytical standard
Sigma-Aldrich
Acetic acid-12C2, 99.9 atom % 12C
Sigma-Aldrich
Sodium bicarbonate, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E500, 99.0-100.5%, powder
SAFC
HEPES
Supelco
DL-Dithiothreitol solution, 1 M in H2O