Open AccessN‐Acetyl‐ d ‐glucosamine‐specific lectin purified from Vibrio cholerae 01
Author(s) -
Sasmal D.,
Guhathakurta B.,
Ghosh A.N.,
Pal C.R.,
Datta A.
Publication year - 1992
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1992.tb05517.x
Subject(s) - antiserum , vibrio cholerae , precipitin , hemagglutinin (influenza) , affinity chromatography , glucosamine , gel electrophoresis , microbiology and biotechnology , biology , ouchterlony double immunodiffusion , immunodiffusion , sodium dodecyl sulfate , biochemistry , chemistry , antigen , bacteria , enzyme , genetics , gene
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