Characterization of Purified Hemagglutinin from Non-Fimbriated Prevotella intermedia

Abstract

Previous studies have shown that partially purified hemagglutinin isolated from non-fimbriated Prevotella intermedia strain E18 can be further purified by arginine agarose and gel filtration. We characterized a purified hemagglutinin and examined the range of genera and species that possesses a common antigen for anti-purified hemagglutinin antiserum.
      Hemagglutination activity was sensitive to heat and enzymes such as trypsin, chymotrypsin, protease, hyaluronidase, lysozyme, β-galactosidase and β-glucosidase. Hemagglutination activity of the hemagglutinin was eliminated by antihemagglutinin antiserum. Residual activity of the hemagglutinin was 50% by addition of galactose and melibiose. Addition of L-arginine and lactose caused hemagglutination inhibition. These results suggest that a common component in the above sugars and amino acid may be associated with receptors for hemagglutinin. Lowering the pH of the hemagglutinin decreased the activity, which reached a residual level of 6.3% at pH2.0.
      Many Prevotella intermedia and a few Prevotella nigrescens strains tested possessed antigen specific to anti-Prevotella intermedia strain E18 hemagglutinin antiserum. In particular, Prevotella intermedia strain 17 cells reacted with the antiserum, indicating that this strain has fimbrial and non-fimbrial hemagglutinins. However, no common antigen was recognized for other species of black-pigmented bacteria. Protein A-gold labeling of Prevotella intermedia strain E18 cells with anti-hemagglutinin IgG revealed that the IgG bound specifically to hemagglutinin present on the Prevotella intermedia strain E18 cell surface, but not on cells that did not react with anti-Prevotella intermedia strain E18 hemagglutinin antiserum.
      These results indicate that the non-fimbriated hemagglutinin in this study is glycoprotein in nature and species-specific.