ABSTRACT
Bacterial infections are the cause of different severe health conditions and new therapies to combat these pathogens have been widely investigated. Carbohydrates, being complex structures covering the surface of bacteria, are considered relevant targets for antibody and vaccine development. The biological activities in pathogenesis of bacterial capsular polysaccharides and lipopolisaccharides and their unique structures have boosted the study of the minimal antigenic binding epitopes and the structural details of antibody-carbohydrate recognition. This review describes the most recent advances on the field, examining the structure, conformation and dynamics of relevant bacterial carbohydrates and their complexes with antibodies. The understanding of key factors governing the recognition process is fundamental for the progress toward the development of specific and efficient bacterial therapeutics.
Subject(s)
Antibodies, Bacterial/immunology , Bacterial Infections/immunology , Bacterial Vaccines/administration & dosage , Drug Development/trends , Polysaccharides, Bacterial/immunology , Antibodies, Bacterial/metabolism , Antibodies, Bacterial/ultrastructure , Bacterial Infections/microbiology , Bacterial Infections/prevention & control , Bacterial Vaccines/immunology , Binding Sites, Antibody/immunology , Drug Development/methods , Epitope Mapping , Epitopes/chemistry , Epitopes/immunology , Epitopes/metabolism , Humans , Molecular Dynamics Simulation , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/metabolism , Protein Structure, Tertiary , Vaccines, Subunit/administration & dosage , Vaccines, Subunit/immunologyABSTRACT
We are confronted daily to unknown microorganisms that have yet to be characterized, detected, and/or analyzed. We propose, in this study, a multidimensional strategy using polyclonal antibodies, consisting of a novel proteomic tool, the ProteomeLab PF2D, coupled to immunological techniques and mass spectrometry (i-PF2D-MS/MS). To evaluate this strategy, we have applied it to Bacillus subtilis, considered here as our unknown bacterial model.
Subject(s)
Antigens, Bacterial/analysis , Bacillus subtilis/immunology , Mass Spectrometry/methods , Proteomics/methods , Amino Acid Sequence , Antibodies, Bacterial/immunology , Antibodies, Bacterial/ultrastructure , Bacillus subtilis/ultrastructure , Blotting, Western , Hydrophobic and Hydrophilic Interactions , Immune Sera , Isoelectric Point , Microscopy, Electron , Molecular Sequence DataABSTRACT
OBJECTIVE: The purpose of this study was to determine the accessibility of monoclonal antibody (McAb), specific for the major cell surface protein antigen (PAg) of Streptococcus sobrinus, to the surface of its native epitopes. MATERIALS AND METHODS: An indirect immunogold labeling technique was used to detect the reaction of McAb with S. sobrinus 6715. The reactions of polyclonal antibodies (PcAbs) against S. sobrinus 6715 or PAg with S. sobrinus 6715, S. mutans Ingbritt C and S. rattus BHT were studied as controls. RESULTS: The results indicated that PAg was localized on the outer cell surface of S. sobrinus, and McAb was reactive with only a few epitopes of the cell surface, whereas PcAbs were found to be reactive with more epitopes. CONCLUSIONS: McAb was specific for the PAg, but there was cross-reaction with S. mutans. Also there seemed to be an association between the fuzzy coat on the surface of S. sobrinus and PAg.
Subject(s)
Adhesins, Bacterial/isolation & purification , Antibodies, Monoclonal/ultrastructure , Bacterial Proteins/isolation & purification , Membrane Glycoproteins , Streptococcus sobrinus/immunology , Animals , Antibodies, Bacterial/isolation & purification , Antibodies, Bacterial/ultrastructure , Bacterial Adhesion/physiology , Microscopy, Immunoelectron , Rats , Streptococcus sobrinus/chemistryABSTRACT
In many prokaryotic cells (eubacteria and archaebacteria) the outermost cell envelope component is composed of a regularly structured protein surface layer (S-layer). The two-dimensional S-layer from Bacillus coagulans E38-66 and Bacillus sphaericus CCM2177 has been investigated by SFM at molecular resolution under physiological conditions (i.e., in buffer solution). We find the E38-66 S-layer lattice to be oblique with lattice parameters of a = 9-10 nm, b = 7-8 nm and gamma = 80 degrees -90 degrees (E38-66). The CCM2177 lattice is square with a = 12-14 nm, in good agreement with TEM data. We have used the unique possibility of the SFM to study the kinematics of biological processes and have performed experiments on the adhesion of polyclonal antibodies to the recrystallized E38-66 protein layer on a time scale of about two to ten seconds per image frame. This represents a first step in directly visualizing molecular recognition reactions.
