Biochemical, immunological and ultrastructural characterization of aggregation substances encoded by Enterococcus faeclis sex‐pheromone plasmids

The sex‐pheromone system of Enterococcus faecalis can be viewed as a unique and highly efficient plasmid‐collection mechanism. The contact needed for transfer of the conjugative sexpheromone plasmids is mediated by an adhesin, called aggregation substance, which is encoded by these plasmids. We show here that for 17 of the 18 sex‐pheromone plasmids (pAM373 being the exception) described to date, their adhesins are immunologically related to each other. In each case, we observed the presence of an N‐terminal fragment of about 78 kDa in addition to the 137‐kDa form of mature aggregation substance. The cross‐reactions were different for the various plasmids. In the case of pPD1 the 78‐kDa fragment reacted only weakly. The aggregation substance encoded by sex‐pheromone plasmid pAD1 (Asa1) was characterized in detail. The conditions used for SDS/PAGE had a drastic influence on the migration behavior of mature aggregation substance and differently migrating, interconvertible forms were identified. Preliminary data indicate that Asa1 might be a glycoprotein. Antibodies were isolated which are directed against the N‐ and C‐terminal parts of aggregation substance. They showed about the same reactivity on Western biots; however, only antibodies directed against the N‐terminal part of the aggregation substance could inhibit the bacterial cell/cell contact. The reactions of the two antibody preparations with induced cells of E.faecalis was analyzed by transmission electron microscopy. The results indicated that especially the N‐terminal part of aggregation substance is exposed on the cell surface of E. faecalis; the C‐terminal part seems to be much less exposed.