Purification of a cell-associated hemagglutinin from Shigella dysenteriae type 1.
A cell-associated hemagglutinin (HA) was isolated and purified from a clinical isolate of Shigella dysenteriae type 1 by affinity chromatography on a fetuin-agarose column. The purified hemagglutinin produced a single-stained protein band of around 66 kDa in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). In an immunodiffusion test, HA-antisera produced a single precipitin band against the purified HA without exhibiting any reactivity towards lipopolysaccharide (LPS) of S. dysenteriae type 1 strain. Inhibition of the hemagglutination by the glycoproteins fetuin, asialofetuin and a sugar derivative N-acetyl-neuraminic acid but not by simple sugars, suggested the specific requirement of complex carbohydrate for binding. Electron micrographs of the purified HA revealed a morphology typical of globular protein.
Guhathakurta
B
B
Division of Biochemistry, National Institute of Cholera and Enteric Diseases, Calcutta, India. icmrnice@ren.nic.in
Sasmal
D
D
Ghosh
A N
AN
Pal
C R
CR
Datta
A
A
D002477:Cells / Q000737:chemistry; D002846:Chromatography, Affinity; D006388:Hemagglutinins / Q000302:isolation & purification* / Q000648:ultrastructure; D012762:Shigella dysenteriae / Q000737:chemistry* / Q000145:classification; D000509:alpha-Fetoproteins
D006388:Hemagglutinins; D000509:alpha-Fetoproteins