Structural Proteins and Glycoproteins of Infectious Bronchitis Virus Particles Labeled during Growth in Chick Embryo Cells

Wadey, C.N.; Westaway, E. G.

doi:10.1159/000149210

Cited by 16 publications

(14 citation statements)

References 20 publications

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“…Competitive ABAs were used for the topographical analysis. The Mr of the proteins precipitated by the various MAbs corresponded to the published Mr of structural proteins of IBV (Cavanagh, 1981;Lomniczi & Morser, 1981 ;Wadey & Westaway, 1981 ;. In addition to a nucleoprotein with an Mr of 49 000, MAb 26.2 precipitated a 45K protein and small amounts of a 39K protein.…”

Section: Discussionmentioning

confidence: 71%

“…The structural proteins of IBV are a phosphorylated nucleoprotein with an Mr of 56000 (56K), a membrane protein that is heterogeneously glycosylated with an Mr ranging from 23000 to 36000 and two glycoproteins, S1 and $2, with an Mr of 92000 and 84000 respectively (Lanser & Howard, 1980;Cavanagh, 1981 ;Lomniczi & Morser, 1981;Wadey & Westaway, 1981;. The spikes or peplomers of IBV are formed from two or three copies each of S 1 and $2 (Cavanagh, 1983a).…”

Section: Introductionmentioning

confidence: 99%

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Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions

Koch

Hartog

Kant

et al. 1990

Journal of General Virology

208

170

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Monoclonal antibodies (MAbs) directed against structural proteins of infectious bronchitis virus (IBV) were produced to analyse the antigenic structure of this virus. Competitive binding of enzyme-labelled and unlabelled MAbs to IBV peplomer protein was analysed in an antibody binding assay to test the relatedness of the epitopes defined by the MAbs. Based on the competition groups, eight epitope clusters were defined (S-A to S-H); six of these clusters (S1-A to S I-F) were located on the S1 subunit and two (S2-G and S2-H) on the $2 subunit of the peplomer protein.Epitope clusters S 1-A and S1-B overlapped extensively. The biological activities of the MAbs were determined and correlated to the epitope clusters. Monoclonal antibodies directed against epitope clusters S 1-A to S1-E and one MAb directed against cluster S2-G moderately to strongly neutralized IBV at titres higher than 2 log~o, whereas the remaining MAbs, directed against S1 and $2, neutralized at titres lower than 2 log~o. One MAb, directed against cluster S1-D, inhibited the agglutination of chicken erythrocytes.

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Section: Discussionmentioning

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions

Koch

Hartog

Kant

et al. 1990

Journal of General Virology

208

170

View full text Add to dashboard Cite

show abstract

“…Nevertheless, the largest glycoproteins of TCV appear to differ fundamentally fi'om those of the well-characterized MHV and IBV viruses. Whereas the largest peplomeric glycoproteins of these two viruses are not changed by the action of reducing agents such as 2-mercaptoethanol [4,32,33,38], the 140,000 tool. wt.-protein of TCV appeared to be reduced to a presumptive 66,000 subunit.…”

Section: Discussionmentioning

confidence: 97%

Identification of the structural proteins of turkey enteric coronavirus

Dea

Tijssen

1988

Archives of Virology

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Coronaviruses in the intestinal contents of turkey poults from Quebec flocks with outbreaks of enteritis were detected by electron microscopy. Five isolates were serially propagated in embryonic turkey eggs by inoculation of clarified intestinal contents into the amniotic cavity. Viral particles contained in the intestinal contents of infected embryos were purified by differential and isopycnic centrifugation in sucrose gradients. Intact virions present in fractions of densities 1.18 to 1.20 g/ml were precipitated with trichloroacetic acid and the protein content was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. At least seven polypeptides with molecular weights ranging from 27,000 to 180,000 were regularly detected after electrophoresis of SDS-solubilized virions. Homologous rabbit antisera to turkey coronaviruses and pooled sera from convalescent turkeys immunochemically stained five polypeptides of apparent molecular weights of 95,000, 72-75,000, 66,000, 52,000 and 27,000. A further polypeptide with a molecular weight of 140,000 appeared in the absence of 2-mercaptoethanol and probably represents a dimer of the 66,000 polypeptide. One Quebec isolate differed from the Minnesota strain by two of its polypeptides, but no antigenic variations could be demonstrated amongst the various isolates either by immuno-electron microscopy, hemagglutination inhibition, or enzyme immuno assays on nitrocellulose.

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“…Two of these, S and M, are glycosylated (Cavanagh, 1981;Lomniczi & Morser, 1981 ;Wadey & Westaway, 1981 ;Stern et al, 1982). My studies of IBV have indicated that the surface projections are associated with two glycopolypeptides, S1 (90K; K = 1000 daltons) and $2 (84K), and that the M protein was associated with two glycopolypeptides of 30K and 28K (Cavanagh, 1981); SI had previously been assigned a mol.…”

Section: Discussionmentioning

confidence: 99%

Coronavirus IBV Glycopolypeptides: Size of Their Polypeptide Moieties and Nature of Their Oligosaccharides

Cavanagh¹

1983

Journal of General Virology

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Analysis of differentially radiolabelled avian infectious bronchitis virus (IBV) indicated that the matrix (M) polypeptides of tool. wt. 23 × 103 (23K), 26K, 28K, 30K and 34K (M23 to M34) which have been shown to give the same peptide maps, differed in their degree of glycosylation; M23 was not glycos'ylated while glycosylation increased with increasing mol. wt. from M26 to M34. Both glucosaminc and mannose were components of M26 to M34 but [3 H]fucose appeared to be associated mainly with M34. Endo-/~-N-acetylglucosaminidase H removed oligosaccharides from M28 and M30 but not M26 and M34, to give a polypeptide of 23K. The surface projection glycopolypeptides SI (90K) and $2 (84K) incorporated 3H-labelled glucosamine and mannosc but not fucose and had oligosaccharides removed by endoglycosidase H. The tool. wt. of the resultant polypeptides varied among experiments; the lowest mol. wt. observed were 64K and 61K. These results indicate (i) that the polypeptide moieties of the S polypcptides are approximately 64K and 61 K, and 23K for the M polypeptide, (ii) that the oligosaccharides of the S and M polypeptides are of the high-mannose type and are linked to the polypeptides by N-glycosidic linkages, and (iii) that the M glycoprotein of IBV differs from that of murinc coronaviruscs and bovine coronavirus L9 which have O-linked oligosaccharides. Avian infectious bronchitis virus (IBV) appears to have three protein structural elements: surface projections (S), a nucleocapsid (N) protein and a matrix/membrane (M) protein (Cavanagh, 1981). Two of these, S and M, are glycosylated (Cavanagh, 1981; Lomniczi & Morser, 1981 ; Wadey & Westaway, 1981 ; Stern et al., 1982). My studies of IBV have indicated that the surface projections are associated with two glycopolypeptides, S1 (90K; K = 1000 daltons) and $2 (84K), and that the M protein was associated with two glycopolypeptides of 30K and 28K (Cavanagh, 1981); SI had previously been assigned a mol. wt. of 94K. Stern et al., (1982) have shown that several polypeptides of IBV, analogous to polypeptides of 34K, 30K/28K, 26K and 23K (M34 to M23) in our studies, all have the same peptide maps. Little is known about the oligosaccharides of the S and M polypeptides. In contrast, it is known from studies with tunicamycin, that the S (or E2) polypeptide(s) of murine and bovine L9 coronaviruses have N-glycosidicaUy linked oligosaccharides while the M (or E 1) glycoprotein has O-glycosidically linked oligosaccharides (Niemann & Klenk, 1981; Holmes et al., 1981; Rottier et al., 1981 ; Siddell et al., 1981 a; Cheley & Anderson, 1981). The glycosylation of the IBV glycopolypeptides has been studied to determine the nature of their oligosaccharides and the mol. wt. of their polypeptide moieties. Working stocks of IBV-M41 and IBV-D41 (Darbyshire et al., 1979) were prepared in embryonated chicken eggs (Cavanagh, 1981). Virus was radiolabelled in pairs of de-embryonated chicken eggs (Cavanagh, 1981). Each egg received 125 ktCi [35S]methionine (sp. act. > 800 Ci/mmol) and one of the following...

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Structural Proteins and Glycoproteins of Infectious Bronchitis Virus Particles Labeled during Growth in Chick Embryo Cells

Cited by 16 publications

References 20 publications

Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions

Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions

Identification of the structural proteins of turkey enteric coronavirus

Coronavirus IBV Glycopolypeptides: Size of Their Polypeptide Moieties and Nature of Their Oligosaccharides

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