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Fractionation and partial characterization of the hemagglutinating and bacterial aggregating adhesins of Bacteroides gingivalis

In order to characterize the surface components responsible for hemagglutination and bacterial aggregation, the outer membrane complex of Bacteroides gingivalis W12 was isolated. It was found to contain both hemagglutinating and bacterial aggregating activity. Examination of the membrane material by biochemical analysis, SDS-polyacrylamide gel electrophoresis and immunological means revealed that the crude outer membrane preparation contained three major proteins and a lipopolysaccharide (LPS) population which displayed size heterogeneity. At least two membrane proteins as well as the LPS were found to be antigenically active using immun-blot analysis. Using gel chromatography and an LPS disaggregating buffer the membrane material was separated into two fractions. An accompanying separation of the two adherenceactivities was observed. The fir s t membrane fraction, containing mostly protein and carbohydrate material was found to contain the bacterial aggregating activity. This fraction also contained a"smooth" IPS population. The second membrane fraction consisted of "rough"type LPS, protein and loosely bound li p i d and was found to contain the hemagglutinating activity. Further investigation revealed that bacterial aggregating activity was blocked by the presence of D-galactose (14 mM) , D-glucosamine (23 mm) and N-acetylglucoscmine (23 mm) and by dialysed iitmune rabbit serum. Dialysed non-immune rabbit serum did not block this activity. Bacterial