Glycosylation of gp116 and gp64 envelope proteins of yellow head virus of Penaeus monodon shrimp.
Yellow head virus (YHV) is a highly virulent pathogen of Penaeus monodon shrimp that is classified in the genus Okavirus, family Roniviridae, in the order Nidovirales. Separation of virion proteins treated with peptide-N-glycosidase-F (PNGase-F) in SDS-polyacrylamide gels and the use of glycoprotein-specific staining methods indicated that the gp116 and gp64 envelope glycoproteins possess N-linked rather than O-linked glycans. Competitive binding inhibition of lectins with various oligosaccharide specificities indicated that glycans linked to gp64 are mannose-rich, whilst glycans linked to gp116 possess terminal N-acetylgalactosamine and N-acetylglucosamine in addition to terminal mannose-type sugars. Mass spectrometry analyses of peptides generated from YHV proteins before and after deglycosylation with PNGase-F, using combinations of the endoproteinases trypsin, Asp-N and Lys-C, confirmed occupancy of six of the seven potential N-linked glycosylation sites in gp116 and three of the four potential sites in gp64.
Soowannayan
Chumporn
C
CSIRO Livestock Industries, Queensland Bioscience Precinct, St Lucia, QLD 4067, Australia. chumporn@biotec.or.th
Cowley
Jeff A
JA
Pearson
Roger D
RD
Wallis
Tristan P
TP
Gorman
Jeffrey J
JJ
Michalski
Wojtek P
WP
Walker
Peter J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000821:virology*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D046828:Roniviridae / Q000737:chemistry / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods; D014759:Viral Envelope Proteins / Q000302:isolation & purification / Q000378:metabolism*
D014759:Viral Envelope Proteins; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase