Lectin-binding epitopes at the surface of Escherichia coli K-12: examination by electron microscopy, with special reference to the presence of a colanic acid-like polymer.
The presence and distribution of lectin-binding epitopes at the surface of Escherichia coli K-12, strain W1655, was studied by electron microscopy after lectin-gold labeling and negative staining. A comparison was made between the lectin-binding capacity of cells cultivated at 20 degrees C and 37 degrees C (in broth or on agar). A variety of pre-treatment protocols were applied prior to labeling. The gold-conjugated lectins used were wheat germ agglutinin (WGA), soybean agglutinin (SBA) and Ulex europaeus lectin (UEA-I). For all culture conditions, the bacteria had moderate exposure of WGA-binding sites, and this was not changed after pre-treatment. Cells cultivated at 37 degrees C had exposed SBA- and UEA-I-binding epitopes apparently associated with the cell surface. These significantly increased in number after boiling the cells for 10 min. With bacteria cultivated at 20 degrees C these two lectins recognized sites situated on exopolysaccharide filaments. Affino dot-blot experiments with isolated polysaccharides of the strain identified the K-12 lipooligosaccharide as the source of WGA-binding epitopes, and the exopolysaccharide, colanic acid (CA) as the source of SBA- and UEA-I-binding sites. The interaction with these two lectins of bacteria cultivated at 37 degrees C could be due to altered translocation of CA from the cytoplasm to the environment. This suggestion was supported by the demonstration by electron microscopy of SBA and UEA-I binding at the surface of hot phenol-water extracted cell walls.
Stoitsova
Stoyanka
S
Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 26, 1113 Sofia, Bulgaria. stoicova@hotmail.com
Ivanova
Radka
R
Dimova
Ivanka
I
D000939:Epitopes / Q000648:ultrastructure*; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D017979:Gold Colloid / Q000737:chemistry; D008070:Lipopolysaccharides; D008854:Microscopy, Electron; D037121:Plant Lectins / Q000737:chemistry / Q000378:metabolism*; D011134:Polysaccharides; D030262:Soybean Proteins / Q000378:metabolism; D013696:Temperature; D014909:Wheat Germ Agglutinins / Q000378:metabolism; D010835:Phytohemagglutinins
D000939:Epitopes; D017979:Gold Colloid; D008070:Lipopolysaccharides; D037121:Plant Lectins; D011134:Polysaccharides; D030262:Soybean Proteins; D014909:Wheat Germ Agglutinins; C036850:Ulex europaeus lectins; C023023:lipid-linked oligosaccharides; C028824:soybean lectin; D010835:Phytohemagglutinins; C004275:colanic acid