Peptide mapping of a novel discontinuous epitope of the major surface adhesin from Streptococcus mutans.
Guy's 13 is a mouse monoclonal antibody that specifically recognizes the major cell-surface adhesion protein SA I/II of Streptococcus mutans, one of the major causative agents of dental caries. Passive immunization with Guy's 13 prevents bacterial colonization in humans. To help elucidate the mechanism of prevention of colonization conferred by this antibody, the SA I/II epitope recognized by Guy's 13 was investigated. It was previously established that the epitope is conformational, being assembled from two non-contiguous regions of SA I/II. In the current study, using recombinant fragments of SA I/II and, ultimately, synthetic peptides, the discontinuous epitope was localized to residues 170-218 and 956-969. This work describes the mapping of a novel discontinuous epitope that requires an interaction between each determinant in order for epitope assembly and recognition by antibody to take place. Guy's 13 binds to the assembled epitope but not to these individual epitope fragments. The assembled epitope results from the interaction between the individual antigenic determinants and can be formed by mixing together determinants present on separate polypeptide chains. The data are consistent with one of the epitope fragments adopting a polyproline II-like helical conformation.
van Dolleweerd
Craig J
CJ
Department of Oral Medicine and Pathology at Guy's, King's and St. Thomas' Hospital Medical Schools, Floor 28 Guy's Tower, Guy's Hospital, London SE1 9RT, United Kingdom. craig.van_dolleweerd@kcl.ac.uk
Kelly
Charles G
CG
Chargelegue
Daniel
D
Ma
Julian K-C
JK
D018829:Adhesins, Bacterial / Q000737:chemistry*; D000409:Alanine / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001665:Binding Sites; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D017353:Gene Deletion; D007074:Immunoglobulin G; D016296:Mutagenesis; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013295:Streptococcus mutans / Q000378:metabolism*
D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D010455:Peptides; D011994:Recombinant Proteins; D000409:Alanine