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Recombinant Soluble Respiratory Syncytial Virus F Protein That Lacks Heptad Repeat B, Contains a GCN4 Trimerization Motif and Is Not Cleaved Displays Prefusion-Like Characteristics.
- Source :
-
PloS one [PLoS One] 2015 Jun 24; Vol. 10 (6), pp. e0130829. Date of Electronic Publication: 2015 Jun 24 (Print Publication: 2015). - Publication Year :
- 2015
-
Abstract
- The respiratory syncytial virus (RSV) fusion protein F is considered an attractive vaccine candidate especially in its prefusion conformation. We studied whether recombinant soluble RSV F proteins could be stabilized in a prefusion-like conformation by mutation of heptad repeat B (HRB). The results show that soluble, trimeric, non-cleaved RSV F protein, produced by expression of the furin cleavage site-mutated F ectodomain extended with a GCN4 trimerization sequence, is efficiently recognized by pre- as well as postfusion-specific antibodies. In contrast, a similar F protein completely lacking HRB displayed high reactivity with prefusion-specific antibodies recognizing antigenic site Ø, but did not expose postfusion-specific antigenic site I, in agreement with this protein maintaining a prefusion-like conformation. These features were dependent on the presence of the GCN4 trimerization domain. Absence of cleavage also contributed to binding of prefusion-specific antibodies. Similar antibody reactivity profiles were observed when the prefusion form of F was stabilized by the introduction of cysteine pairs in HRB. To study whether the inability to form the 6HB was responsible for the prefusion-like antibody reactivity profile, alanine mutations were introduced in HRB. Although introduction of alanine residues in HRB inhibited the formation of the 6HB, the exposure of postfusion-specific antigenic site I was not prevented. In conclusion, proteins that are not able to form the 6HB, due to mutation of HRB, may still display postfusion-specific antigenic site I. Replacement of HRB by the GCN4 trimerization domain in a non-cleaved soluble F protein resulted, however, in a protein with prefusion-like characteristics, suggesting that this HRB-lacking protein may represent a potential prefusion F protein subunit vaccine candidate.
- Subjects :
- Antibodies, Neutralizing pharmacology
Binding Sites
Cell Line, Tumor
Epithelial Cells pathology
Epithelial Cells virology
Gene Expression
HEK293 Cells
Humans
Models, Molecular
Protein Binding
Protein Multimerization
Protein Structure, Tertiary
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Respiratory Mucosa pathology
Respiratory Mucosa virology
Respiratory Syncytial Virus, Human metabolism
Viral Fusion Proteins antagonists & inhibitors
Viral Fusion Proteins chemistry
Viral Fusion Proteins metabolism
Antibodies, Viral pharmacology
Epithelial Cells metabolism
Respiratory Mucosa metabolism
Respiratory Syncytial Virus, Human genetics
Viral Fusion Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 10
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 26107504
- Full Text :
- https://doi.org/10.1371/journal.pone.0130829