Saliva-mediated aggregation of Enterococcus faecalis transformed with a Streptococcus sanguis gene encoding the SSP-5 surface antigen.
The interaction of a high-molecular-weight salivary glycoprotein (agglutinin) with Streptococcus sanguis M5 leads to the formation of bacterial aggregates. We have previously shown that the SSP-5 surface antigen from S. sanguis M5 binds the salivary agglutinin and therefore may be involved in the aggregation process. Here we report the transformation of a nonaggregating Enterococcus faecalis strain with the SSP-5 gene and show that the protein is expressed on the cell surface and confers an aggregation-positive phenotype. E. faecalis S161 protoplasts were transformed with pAM401 EB-5, a shuttle vector containing the S. sanguis SSP-5 gene, resulting in the isolation of E. faecalis S161EB-5. Crude cell extracts from this transformant and from S. sanguis M5 were analyzed by Western blotting. Extracts from S. sanguis M5 possessed peptides of 190 and 205 kilodaltons that reacted strongly with polyclonal antibodies against the recombinant SSP-5 antigen. E. faecalis S161EB-5 contained only the 190-kilodalton immunoreactive protein, suggesting that the antigen may be processed differently in E. faecalis S161EB-5. The parent strain, E. faecalis S161, did not react with this antibody preparation. Immunogold labeling of intact E. faecalis S161EB-5 and S. sanguis M5 with anti-SSP-5 immunoglobulin G showed that both organisms expressed similar levels of the antigen. Both organisms formed visible aggregates upon incubation with salivary agglutinin. These results suggest that the SSP-5 antigen may mediate both the binding of agglutinin to S. sanguis M5 and the subsequent formation of bacterial aggregates.
Demuth
D R
DR
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104.
Berthold
P
P
Leboy
P S
PS
Golub
E E
EE
Davis
C A
CA
Malamud
D
D
D000373:Agglutinins / Q000494:pharmacology; D000942:Antigens, Bacterial / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics*; D004755:Enterobacteriaceae / Q000235:genetics; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques; D011994:Recombinant Proteins / Q000276:immunology; D012463:Saliva / Q000382:microbiology*; D013298:Streptococcus sanguis / Q000235:genetics / Q000276:immunology*; D014170:Transformation, Genetic
D000373:Agglutinins; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D011994:Recombinant Proteins