4HEP

Complex of lactococcal phage TP901-1 with a llama vHH (vHH17) binder (nanobody)

Summary for 4HEP

• Entry DOI: 10.2210/pdb4hep/pdb
• Related: 3EJC 3F0C 3HG0 4HEM 4IOS
• Descriptor: BPP, vHH17 domain, SULFATE ION, ... (4 entities in total)
• Functional Keywords: alpha-beta, phage receptor binding protein, llama glama vhh domain, viral protein, cell adhesion-immune system complex
• Biological source: Lactococcus phage TP901-1; More
• Total number of polymer chains: 2
• Total formula weight: 31287.78

Authors

Desmyter, A.,Spinelli, S.,Farenc, C.,Blangy, S.,Bebeacua, C.,van Sinderen, D.,Mahony, J.,Cambillau, C. (deposition date: 2012-10-04, release date: 2013-03-20, Last modification date: 2024-10-30)

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 (1.75 Å)