Detection of the ganglioside N-glycolyl-neuraminyl-lactosyl-ceramide by biotinylated Escherichia coli K99 lectin.
K99 lectin from Escherichia coli was purified and biotinylated via its carboxyl groups using biocytin hydrazide and a water soluble carbodiimide. Biotinylation of two out of the nine carboxyl groups was sufficient to permit detection of the lectin by avidin and did not cause any loss of the haemagglutinating activity. It was demonstrated that the biotinylated K99 lectin retained other important properties of native K99 and that it will probably become a very sensitive detecting reagent. Indeed, it was able to bind to HeLa cells, as do intact bacteria carrying K99 fimbriae, and also to recognize N-glycolyl-neuraminyl-lactosyl-ceramide in an overlay binding assay.
Ouadia
A
A
Institut de Biotechnologie, Université de Limoges, France.
Karamanos
Y
Y
Julien
R
R
D000954:Antigens, Surface / Q000302:isolation & purification; D001422:Bacterial Adhesion / Q000502:physiology; D001427:Bacterial Toxins; D001710:Biotin; D002240:Carbohydrate Sequence; D002264:Carboxylic Acids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005679:G(M3) Ganglioside / Q000031:analogs & derivatives* / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data
D000954:Antigens, Surface; D001427:Bacterial Toxins; D002264:Carboxylic Acids; D005679:G(M3) Ganglioside; C031429:K99 antigen; D008562:Membrane Glycoproteins; C044298:N-glycolylneuraminyllactosylceramide; D001710:Biotin