title: Identification and analysis of gp116 and gp64 structural glycoproteins of yellow head nidovirus of Penaeus monodon shrimp.
abstract: Yellow head virus (YHV) is a major agent of disease in farmed penaeid shrimp. YHV virions purified from infected shrimp contain three major structural proteins of molecular mass 116 kDa (gp116), 64 kDa (gp64) and 20 kDa (p20). Two different staining methods indicated that the gp116 and gp64 proteins are glycosylated. Here we report the complete nucleotide sequence of ORF3, which encodes a polypeptide of 1666 amino acids with a calculated molecular mass of 185 713 Da (pI=6.68). Hydropathy analysis of the deduced ORF3 protein sequence identified six potential transmembrane helices and three ectodomains containing multiple sites for potential N-linked and O-linked glycosylation. N-terminal sequence analysis of mature gp116 and gp64 proteins indicated that each was derived from ORF3 by proteolytic cleavage of the polyprotein between residues Ala(228) and Thr(229), and Ala(1127) and Leu(1128), located at the C-terminal side of transmembrane helices 3 and 5, respectively. Comparison with the deduced ORF3 protein sequence of Australian gill-associated virus (GAV) indicated 83 % amino acid identity in gp64 and 71 % identity in gp116, which featured two significant sequence deletions near the N terminus. Database searches revealed no significant homology with other proteins. Recombinant gp64 expressed in E. coli with and without the C-terminal transmembrane region was shown to react with antibody raised against native gp64 purified from virions.
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.|CENTEX Shrimp, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Sarawut
authors.initials: S
authors.lastname: Jitrapakdee
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.|CENTEX Shrimp, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Sasimanas
authors.initials: S
authors.lastname: Unajak
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.|CENTEX Shrimp, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Nusra
authors.initials: N
authors.lastname: Sittidilokratna
authors.affiliation: CSIRO Livestock Industries, Long Pocket Laboratories, Indooroopilly, Queensland, Australia.
authors.forename: Richard A J
authors.initials: RAJ
authors.lastname: Hodgson
authors.affiliation: CSIRO Livestock Industries, Long Pocket Laboratories, Indooroopilly, Queensland, Australia.
authors.forename: Jeff A
authors.initials: JA
authors.lastname: Cowley
authors.affiliation: CSIRO Livestock Industries, Long Pocket Laboratories, Indooroopilly, Queensland, Australia.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Walker
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Sakol
authors.initials: S
authors.lastname: Panyim
authors.affiliation: Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.|CENTEX Shrimp, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Vichai
authors.initials: V
authors.lastname: Boonsaeng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004926:Escherichia coli / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D006031:Glycosylation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D028381:Nidovirales / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016366:Open Reading Frames; D033561:Penaeidae / Q000821:virology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013194:Staining and Labeling; D015678:Viral Structural Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D006023:Glycoproteins; D011994:Recombinant Proteins; D015678:Viral Structural Proteins