Open AccessPurification of a mannose/glucose‐specific hemagglutinin/lectin from a Vibrio cholerae O1 strain
Author(s) -
Sasmal D,
Guhathakurta B,
Ghosh A.N,
Pal C.R,
Datta A
Publication year - 1999
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.1999.tb01242.x
Subject(s) - vibrio cholerae , antiserum , biology , hemagglutinin (influenza) , microbiology and biotechnology , mannose , affinity chromatography , lectin , vibrionaceae , biochemistry , vibrio , bacteria , antigen , genetics , gene , enzyme
A cell‐associated mannose/glucose‐specific hemagglutinin (MSHA) has been purified from a strain of Vibrio cholerae O1 by chromatography on a chitin column followed by affinity purification on Sephadex G100. The purified protein gave a single stained band of 40 kDa by SDS‐PAGE, exhibited high affinity towards d ‐mannose and d ‐glucose but was resistant to l ‐fucose and N ‐acetyl‐ d ‐glucosamine. The purified MSHA was revealed as a globular form of protein under electron microscope. In immunodiffusion tests the purified MSHA produced a single precipitin band against homologous antisera and antisera raised against the whole cell bacteria without any reactivity towards antisera raised against the purified N ‐acetyl‐ d ‐glucosamine‐specific lectin of the same bacterial strain. Immunogold labelling confirmed the location of hemagglutinin on the surface of the bacteria. Purified MSHA reacted strongly with sera from convalescent cholera patients in immunodiffusion tests.