N-Acetyl-d-glucosamine-specific lectin purified fromVibrio cholerae01

Abstract

An N‐acetyl‐d‐glucosamine‐specific cell associated hemagglutinin (HA) was isolated and purified from a strain of Vibrio cholerae 01 by chitin affinity chromatography followed by separation on Bio Gel P‐150. A single stained protein band of 47 kDa in sodium dodecyl sulfate‐polyacryl‐amide gel electrophoresis (SDS‐PAGE) was observed with the purified HA. HA‐antisera produced a single precipitin band against the purified HA in an immunodiffusion test without exhibiting any reactivity towards purified lipopolysaccharide (LPS). Purified HA, used as solid‐phase antigen in an enzyme‐linked immunosorbent assay (ELISA), reacted strongly with HA‐antisera but cross‐reacted negligibly with antisera raised against purified LPS. Hemagglutinating activity of the purified HA was highly sensitive to N‐acetyl‐d‐glucosamine. The immunogold‐labelling method using HA‐antisera confirmed the location of the HA on the surface of the bacterial cells. The HA‐antisera reacted with

Studies on adhesion, haemagglutination and other biological properties ofVibrio choleraeO139

“…The K cholerae 0139 strains did not react immunologically with the purified CAHA from V. cholerae 01, which had been suggested as an adhesin [3]. The moderately high cell-associated haemagglutinating activity and high adhesive capacity to RIEC are therefore due to some other molecular entity unrelated to the purified CAHA from V: cholerae 01.…”

“…The purified cell-associated haemagglutinin (CAHA) from I! cholerue 01 acted as an adhesin molecule in an in vitro system with isolated RIEC [3]. Among the other pathogenic fac- tors a toxin co-regulated pilus designated as TCP in pathogenic I/; cholerue 01 was involved in the colonization of the human intestine [21.…”

N-Acetyl-?-glucosamine specific hemagglutinin receptor of Vibrio cholerae O1 in chicken erythrocyte membranes

“…Vibrio cholerae O1 is known to produce soluble and cellassociated hemagglutinin with di¡erent sugar speci¢cities [2^5]. A cell-associated N-acetyl-D-glucosamine speci¢c hemagglutinin (HA) was isolated and puri¢ed from a strain of Vibrio cholerae O1 by a¤nity chromatography on a chitin column followed by BioGel ¢ltration [6]. The puri¢ed HA was documented as an adhesin molecule in V. cholerae O1 cells as studied in adhesion assays using isolated rabbit intestinal epithelial cells (RIEC) and rabbit intestinal brush borders (RIBB) in the presence of HA antisera and Fab (IgG) fragments of the HA antibody [7].…”

Studies on adhesion, haemagglutination and other biological properties of Vibrio cholerae O139

“…The purified cell-associated haemagglutinin (CAHA) from I! cholerue 01 acted as an adhesin molecule in an in vitro system with isolated RIEC [3]. Among the other pathogenic fac-200 tors a toxin co-regulated pilus designated as TCP in pathogenic I/; cholerue 01 was involved in the colonization of the human intestine [21.…”