N-acetyl-D-glucosamine-specific lectin purified from Vibrio cholerae 01.
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-polyacrylamide 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 a protein component of the homologous outer membrane preparation. A significant inhibition was observed in the adhesive capability of the V. cholerae 01 strain to isolated rabbit intestinal epithelial cells (RIEC) in vitro when the later were pre-treated with the purified HA.
Sasmal
D
D
National Institute of Cholera and Enteric Diseases, Calcutta, India.
Guhathakurta
B
B
Ghosh
A N
AN
Pal
C R
CR
Datta
A
A
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D006388:Hemagglutinins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007422:Intestines / Q000382:microbiology; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D011817:Rabbits; D014734:Vibrio cholerae / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D006388:Hemagglutinins; D037102:Lectins; D000117:Acetylglucosamine