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J. Biol. Chem., Vol. 279, Issue 21, 22198-22203, May 21, 2004

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Peptide Mapping of a Novel Discontinuous Epitope of the Major Surface Adhesin from Streptococcus mutans^*

Craig J. van Dolleweerd, Charles G. Kelly, Daniel Chargelegue¶, and Julian K-C. Ma¶

From the 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 and the ^¶Department of Infectious Diseases at St. George's Hospital Medical School, London SW17 ORE, United Kingdom

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.

Received for publication, January 26, 2004

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 44-207-188-43-57; Fax: 44-207-188-43-75; E-mail: craig.van_dolleweerd{at}kcl.ac.uk.

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