Peer-Reviewed Journal Details
Mandatory Fields
Desmyter, A,Farenc, C,Mahony, J,Spinelli, S,Bebeacua, C,Blangy, S,Veesler, D,van Sinderen, D,Cambillau, C
2013
April
Proceedings of The National Academy of Sciences of The United States of America
Viral infection modulation and neutralization by camelid nanobodies
Validated
Optional Fields
LACTOCOCCUS-LACTIS PHAGES RECEPTOR-BINDING PROTEIN ELECTRON-MICROSCOPY MAXIMUM-LIKELIHOOD ANTIBODY FRAGMENT CRYSTAL-STRUCTURE ESCHERICHIA-COLI TAIL BASEPLATE MECHANISM
110
1371
1379
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 Ca2+, 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.
10.1073/pnas.1301336110
Grant Details