Modular Structure of the Receptor Binding Proteins of Lactococcus lactis Phages

Lactococcus lactis is a Gram-positive bacterium widely used by the dairy industry. Several industrial L. lactis strains are sensitive to various distinct bacteriophages. Most of them belong to the Siphoviridae family and comprise several species, among which the 936 and P335 are prominent. Members of these two phage species recognize their hosts through the interaction of their receptor-binding protein (RBP) with external cell wall saccharidices of the host, the “receptors.” We report here the 1.65 Å resolution crystal structure of the RBP from phage TP901-1, a member of the P335 species. This RBP of 163 amino acids is a homotrimer comprising three domains: a helical N terminus, an interlaced β-prism, and a β-barrel, the head domain (residues 64-163), which binds a glycerol molecule. Fluorescence quenching experiments indicated that the RBP exhibits high affinity for glycerol, muramyl-dipeptide, and other saccharides in solution. The structural comparison of this RBP with that of lactococcal phage p2 RBP, a member of the 936 species (Spinelli, S., Desmyter, A., Verrips, C. T., de Haard, J. W., Moineau, S., and Cambillau, C. (2006) Nat. Struct. Mol. Biol. 13, 85-89) suggests a large extent of modularity in RBPs of lactococcal phages.

[1]  A. Desmyter,et al.  Receptor-Binding Protein of Lactococcus lactis Phages: Identification and Characterization of the Saccharide Receptor-Binding Site , 2006, Journal of bacteriology.

[2]  H. Neve,et al.  Identification of the Lower Baseplate Protein as the Antireceptor of the Temperate Lactococcal Bacteriophages TP901-1 and Tuc2009 , 2006, Journal of bacteriology.

[3]  E. Dodson,et al.  Structure of a group A streptococcal phage-encoded virulence factor reveals a catalytically active triple-stranded beta-helix. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Sylvain Moineau,et al.  Llama Antibodies against a Lactococcal Protein Located at the Tip of the Phage Tail Prevent Phage Infection , 2005, Journal of bacteriology.

[5]  H. Neve,et al.  Structural Characterization and Assembly of the Distal Tail Structure of the Temperate Lactococcal Bacteriophage TP901-1 , 2005, Journal of bacteriology.

[6]  P. Su,et al.  Lactococcal 936-species phage attachment to surface of Lactococcus lactis. , 2005, Journal of dairy science.

[7]  G. Gao,et al.  Crystal Structure of Severe Acute Respiratory Syndrome Coronavirus Spike Protein Fusion Core* , 2004, Journal of Biological Chemistry.

[8]  W. Weissenhorn,et al.  Class I and class II viral fusion protein structures reveal similar principles in membrane fusion (Review) , 2004, Molecular membrane biology.

[9]  H. Neve,et al.  Identification of the Receptor-Binding Protein in 936-Species Lactococcal Bacteriophages , 2004, Applied and Environmental Microbiology.

[10]  F. Vogensen,et al.  Identification of Lactococcus lactis Genes Required for Bacteriophage Adsorption , 2004, Applied and Environmental Microbiology.

[11]  S. Cusack,et al.  Crystal Structure of Species D Adenovirus Fiber Knobs and Their Sialic Acid Binding Sites , 2004, Journal of Virology.

[12]  Renaud Vincentelli,et al.  A medium-throughput crystallization approach. , 2002, Acta crystallographica. Section D, Biological crystallography.

[13]  H. Deveau,et al.  Effect of Exopolysaccharides on Phage-Host Interactions in Lactococcus lactis , 2002, Applied and Environmental Microbiology.

[14]  D. Stuart,et al.  Evolution of viral structure. , 2002, Theoretical population biology.

[15]  C. Verrips,et al.  Preventing phage lysis of Lactococcus lactis in cheese production using a neutralizing heavy-chain antibody fragment from llama. , 2002, Journal of dairy science.

[16]  Thilo Stehle,et al.  Crystal structure of reovirus attachment protein σ1 reveals evolutionary relationship to adenovirus fiber , 2002, The EMBO journal.

[17]  S. Miller,et al.  Crystal structure of a heat and protease-stable part of the bacteriophage T4 short tail fibre. , 2001, Journal of molecular biology.

[18]  F. Vogensen,et al.  Analysis of the complete DNA sequence of the temperate bacteriophage TP901-1: evolution, structure, and genome organization of lactococcal bacteriophages. , 2001, Virology.

[19]  H. Deveau,et al.  Characterization of mesophilic mixed starter cultures used for the manufacture of aged cheddar cheese. , 2000, Journal of dairy science.

[20]  Liam J. McGuffin,et al.  The PSIPRED protein structure prediction server , 2000, Bioinform..

[21]  P. Hols,et al.  The biosynthesis and functionality of the cell-wall of lactic acid bacteria , 1999, Antonie van Leeuwenhoek.

[22]  M G Rossmann,et al.  Structure of bacteriophage T4 fibritin: a segmented coiled coil and the role of the C-terminal domain. , 1997, Structure.

[23]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[24]  S. Moineau,et al.  Isolation and Characterization of Lactococcal Bacteriophages from Cultured Buttermilk Plants in the United States , 1996 .

[25]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[26]  J. Navaza,et al.  AMoRe: an automated package for molecular replacement , 1994 .

[27]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[28]  T. Klaenhammer,et al.  Restriction/Modification Systems and Restriction Endonucleases Are More Effective on Lactococcal Bacteriophages That Have Emerged Recently in the Dairy Industry , 1993, Applied and environmental microbiology.

[29]  H. Ackermann,et al.  Characterization of lactococcal bacteriophages from Quebec cheese plants , 1992 .

[30]  W. Sandine,et al.  The bacteriophage kh receptor of Lactococcus lactis subsp. cremoris KH is the rhamnose of the extracellular wall polysaccharide , 1990, Applied and environmental microbiology.

[31]  B. Matthews.  Solvent content of protein crystals. , 1968, Journal of molecular biology.

[32]  Sylvain Moineau,et al.  Lactococcal bacteriophage p2 receptor-binding protein structure suggests a common ancestor gene with bacterial and mammalian viruses , 2006, Nature Structural &Molecular Biology.

[33]  U. Baumann,et al.  An efficient one-step site-directed and site-saturation mutagenesis protocol. , 2004, Nucleic acids research.

[34]  A. Mitraki,et al.  Review: conformation and folding of novel beta-structural elements in viral fiber proteins: the triple beta-spiral and triple beta-helix. , 2002, Journal of structural biology.

[35]  M. Vidal,et al.  GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. , 2000, Methods in enzymology.

[36]  S. Salminen.  Lactic acid bacteria: microbiology and functional aspects. , 1998 .

[37]  W. Sandine,et al.  Lactococcus lactis ssp. lactis C2 Bacteriophage sk1 Receptor involving Rhamnose and Glucose Moieties in the Cell Wall , 1994 .

[38]  E. W. Nielsen,et al.  Plasmid profiles and bacteriophage sensitivity of bacteria of a Cheddar starter used for five years without rotation , 1988 .