Characterization of a hemagglutinin derived from Prevotella nigrescens
Keywords:
Hemagglutinin,
(Prevotella nigrescens)
1996 Volume 59 Issue 2 Pages 127-136
Details
1996 Volume 59 Issue 2 Pages 127-136
We isolated a hemagglutinin from Prevotella nigrescens strain E 4 by mechanical shearing, ultrasonication and gel filtration. Hemagglutinating activity was eluted in the fractions following the first and second peaks on a Sepharose CL-4 B column. The Ouchterlony method revealed that both active fractions react with IgG against a purified hemagglutinin from non-fimbriated Prevotella intermedia strain E 18 (E 18 HA). The precipitating line between first active fraction and anti-E 18 HA IgG was located in the center of the wells, although the line between E 4 HA and anti-E 18 HA IgG was located on the side of the antibody. In a previous report, we disclosed that Prevotella intermedia ATCC 15032 possess a strong hemagglutinating activity, but do not show precipitation with E 18 HA antiserum, while Prevotella nigrescens strains P 43 and P 77 do not show hemagglutinating activity and precipitation with E 18 HA IgG. Therefore, Prevotella intermedia ATCC 15032, Prevotella nigrescens strains P 43 and p 77 cells were harvested, mechanically sheared, sonicated, and reacted with E 18 HA IgG. We found a common antigen between E 18 HA and Prevotella intermedia ATCC 15032 cells, but not in Prevotella nigrescens strains P 43 and P 77. We obtained a single band at approximately 40k Da when we applied the second peak (designated E 4 HA) to SDS-PAGE. Western blotting produced a single band corresponding to those from SDS-PAGE. The activity was sensitive to trypsin, chymotrypsin, protease, β-glucosidase and various detergents, but resistant to heat at 80°C for 10 min. Addition of L-arginine and L-lysine caused hemagglutination inhibition, although addition of L-fucose, L-rhamnose and maltose increased the activity.
These results indicate that this hemagglutinin is a protein-polysaccharide complex in nature and is distributed on most of the cell surface of Prevotella intermedia and Prevotella nigrescens. Shika Igaku (J Osaka Odontol Soc) 1996 Jun; 59(2): 127-136.
