4IOS

Structure of phage TP901-1 RBP (ORF49) in complex with nanobody 11.

Summary for 4IOS

• Entry DOI: 10.2210/pdb4ios/pdb
• Related: 4HEM 4HEP
• Descriptor: BPP, Llama nanobody 11, GLYCEROL, ... (4 entities in total)
• Functional Keywords: all beta, jelly roll motif, receptor binding protein, neutralizing llama antibody domain, cell adhesion-immune system complex, viral protein
• Biological source: Lactococcus phage TP901-1; More
• Total number of polymer chains: 8
• Total formula weight: 95502.13

Authors

Desmyter, A.,Farenc, C.,Mahony, J.,Spinelli, S.,Bebeacua, C.,Blangy, S.,Veesler, D.,van Sinderen, D.,Cambillau, C. (deposition date: 2013-01-08, release date: 2013-03-20, Last modification date: 2024-11-13)

Primary citation

Desmyter, A.,Farenc, C.,Mahony, J.,Spinelli, S.,Bebeacua, C.,Blangy, S.,Veesler, D.,van Sinderen, D.,Cambillau, C.
Viral infection modulation and neutralization by camelid nanobodies
Proc.Natl.Acad.Sci.USA, 110:E1371-E1379, 2013

PubMed 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.

PubMed: 23530214
DOI: 10.1073/pnas.1301336110

Experimental method

X-RAY DIFFRACTION (2.4 Å)