Expression of the F glycoprotein gene from human respiratory syncytial virus inEscherichia coli: Mapping of a fusion inhibiting epitope

Martin-Gallardo, Antonia; Fien, Karen; Hu, Branda; Farley, John F.; Seid, Robert C.; Collins, Peter L.; Hildreth, Stephen W.; Paradiso, Peter R.

doi:10.1016/0042-6822(91)90863-7

Cited by 22 publications

(13 citation statements)

References 16 publications

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“…Martin-Gallardo et al (1991) found that the hydrophobic N-terminal signal sequence was toxic when the eDNA was introduced into a bacterial expression vector under the inducible control of the 2 promoter. A full-length gene product was not detected but three cleavage products were expressed ranging from M r 18 500 to 45 000.…”

Section: Discussionmentioning

confidence: 99%

Binding of neutralizing monoclonal antibodies to regions of the fusion protein of respiratory syncytial virus expressed in Escherichia coli

Lounsbach

Bourgeois

West

et al. 1993

Journal of General Virology

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Section: Discussionmentioning

confidence: 99%

Binding of neutralizing monoclonal antibodies to regions of the fusion protein of respiratory syncytial virus expressed in Escherichia coli

Lounsbach

Bourgeois

West

et al. 1993

Journal of General Virology

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“…Several laboratories have located a few epitopes on the primary structure of the F protein by (i) using synthetic peptides to examine the binding of MAbs (Trudel et al, 1987;Bourgeois et al, 1991;Martin-Gallardo et al, 1991) or polyclonal antibodies (Scopes et al, 1990), or (ii) isolating viruses that escape neutralization by anti-F protein MAbs in order to identify amino acid residues essential for epitope integrity . Since both approaches have limitations (see Discussion), we have used the two complementary strategies to locate the epitopes recognized by several anti-F protein neutralizing MAbs on the primary structure of the protein.…”

Section: Introductionmentioning

confidence: 99%

Characterization of two antigenic sites recognized by neutralizing monoclonal antibodies directed against the fusion glycoprotein of human respiratory syncytial virus

Arbiza

Taylor²,

López

et al. 1992

Journal of General Virology

138

142

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Two antigenic sites recognized by neutralizing monoclonal antibodies (MAbs) directed against the fusion (F) glycoprotein of human respiratory syncytial virus were mapped on the primary structure of the protein by (i) the identification of amino acid substitutions selected in antibody-escape mutants and (ii) the reactivity of synthetic peptides with MAbs. The first site contained several overlapping epitopes which were located within the trypsin-resistant amino-terminal third of the large F1 subunit. Only one of these epitopes was faithfully reproduced by a short synthetic peptide; the others might require specific local conformations to react with MAbs. The second antigenic site was located in a trypsin-sensitive domain of the F1 subunit towards the carboxy-terminal end of the cysteine-rich region. One of these epitopes was reproduced by synthetic peptides. In addition, mutagenized F protein with a substitution of serine for arginine at position 429 did not bind MAbs to the second site. These results are discussed in terms of F protein structure and the mechanisms of virus neutralization.

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“…Other authors have found patterns similar to these. Although the binding sites of a number of neutralizing and fusion-inhibiting antibodies have been mapped to two regions of the protein, involving or close to the amino acids 265 to 272 (Arbiza et al, 1992;Lounsbach et al, 1993;Martin-Gallardo et al, 1991) and amino acids 422 to 438 (Lounsbach et aL, 1993). Each fragment is labelled according to the amino acids of the F protein sequence expressed in the chimera.…”

mentioning

confidence: 99%

“…One of these MAbs, B4, also bound to a synthetic peptide corresponding to residues 255 to 275. This region is contiguous with the binding site of MAb L4, amino acids 289 to 298, also determined by binding to F protein fragments expressed in E. coli (Martin-Gallardo et al, 1991). These binding sites lie in the N-terminal, e helix-rich half of the cytoplasmic domain of the F 1 protein.…”

mentioning

confidence: 99%

Biological activity, binding site and affinity of monoclonal antibodies to the fusion protein of respiratory syncytial virus

West

Lounsbach²,

Bourgeois³

et al. 1994

Journal of General Virology

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The neutralizing activity and fusion-inhibition activity per unit weight of immunoglobulin were determined for each of a panel of 20 monoclonal antibodies (MAbs) to the fusion (F) protein of respiratory syncytial (RS) virus. Neutralization did not correlate with fusion-inhibiting activity, suggesting that the F protein plays at least two independent, antibody-sensitive roles in viral infection. Antibodies with the highest biological activity against A2, a subgroup A strain of RS virus, neutralized a subgroup B strain (8/60) poorly, suggesting a degree of antigenic variation that may be important in human infection.All but one fusion-inhibiting MAb bound to protein blots and binding was mapped to two areas on overlapping F protein fragments. One MAb with relatively poor fusion-inhibiting activity bound only to fragments C-terminal of amino acid 384, the remainder bound only to fragments containing residues 253 to 289. MAbs directed to the latter site were heterogeneous in neutralizing activity, subgroup specificity and fusioninhibiting activity. These variations between MAbs could not be accounted for by differences in their binding avidities. We suggest that this binding site is not the complete antibody epitope which probably includes conformation-dependent elements.

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Expression of the F glycoprotein gene from human respiratory syncytial virus inEscherichia coli: Mapping of a fusion inhibiting epitope

Cited by 22 publications

References 16 publications

Binding of neutralizing monoclonal antibodies to regions of the fusion protein of respiratory syncytial virus expressed in Escherichia coli

Binding of neutralizing monoclonal antibodies to regions of the fusion protein of respiratory syncytial virus expressed in Escherichia coli

Characterization of two antigenic sites recognized by neutralizing monoclonal antibodies directed against the fusion glycoprotein of human respiratory syncytial virus

Biological activity, binding site and affinity of monoclonal antibodies to the fusion protein of respiratory syncytial virus

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