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Viral infection modulation and neutralization by camelid nanobodies
- Source :
- Proceedings of the National Academy of Sciences of the United States of America, Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2013, 110 (15), pp.E1371-E1379. ⟨10.1073/pnas.1301336110⟩, Proceedings of the National Academy of Sciences of the United States of America, 2013, 110 (15), pp.E1371-E1379. ⟨10.1073/pnas.1301336110⟩
- Publication Year :
- 2013
- Publisher :
- HAL CCSD, 2013.
-
Abstract
- Lactococcal phages belong to a large family of Siphoviridae and infect Lactococcus lactis, a gram-positive bacterium used in commercial dairy fermentations. These phages are believed to recognize and bind specifically to pellicle polysaccharides covering the entire bacterium. The phage TP901-1 baseplate, located at the tip of the tail, harbors 18 trimeric receptor binding proteins (RBPs) promoting adhesion to a specific lactococcal strain. Phage TP901-1 adhesion does not require major conformational changes or Ca(2+), which contrasts other lactococcal phages. Here, we produced and characterized llama nanobodies raised against the purified baseplate and the Tal protein of phage TP901-1 as tools to dissect the molecular determinants of phage TP901-1 infection. Using a set of complementary techniques, surface plasmon resonance, EM, and X-ray crystallography in a hybrid approach, we identified binders to the three components of the baseplate, analyzed their affinity for their targets, and determined their epitopes as well as their functional impact on TP901-1 phage infectivity. We determined the X-ray structures of three nanobodies in complex with the RBP. Two of them bind to the saccharide binding site of the RBP and are able to fully neutralize TP901-1 phage infectivity, even after 15 passages. These results provide clear evidence for a practical use of nanobodies in circumventing lactococcal phages viral infection in dairy fermentation.
- Subjects :
- Models, Molecular
Viral protein
viruses
Phagemid
Molecular Conformation
Siphoviridae
Crystallography, X-Ray
medicine.disease_cause
Epitope
Microbiology
Epitopes
03 medical and health sciences
Protein structure
Antibody Specificity
medicine
Animals
Nanotechnology
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
Infectivity
0303 health sciences
Binding Sites
Multidisciplinary
biology
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
030302 biochemistry & molecular biology
Lactococcus lactis
[SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN]
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
Viral Tail Proteins
Single-Domain Antibodies
Surface Plasmon Resonance
biology.organism_classification
Protein Structure, Tertiary
Microscopy, Electron
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
PNAS Plus
Fermentation
[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology
Camelids, New World
Subjects
Details
- Language :
- English
- ISSN :
- 00278424 and 10916490
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America, Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2013, 110 (15), pp.E1371-E1379. ⟨10.1073/pnas.1301336110⟩, Proceedings of the National Academy of Sciences of the United States of America, 2013, 110 (15), pp.E1371-E1379. ⟨10.1073/pnas.1301336110⟩
- Accession number :
- edsair.doi.dedup.....2b83a3b8eaaf5f08044a0e3505df865b