Characterization and use of biotinylated Escherichia coli K99 lectin.
K99 lectin from Escherichia coli was purified and biotinylated via the amino groups of lysine residues using N-biotinyl-6-amino-caproic acid N-hydroxysuccinimide ester (BcapNHS). Biotin was detected on Lys-47 and Lys-87. It was previously demonstrated (Jacobs, A.A.C., Van den Berg, P.A., Bak, H.J. and De Graaf, F.K. (1986) Biochim. Biophys. Acta 872, 92-97) that modification of lysine residues 132 and 133 with 4-chloro-3,5-dinitrobenzoate (CDNB) resulted in the loss of the binding capacity of K99 fimbriae. Due to the higher size of the biotin derivative compared to CDNB, Lys-132 or Lys-133, essential for the biological activity, were not modified. The biotinylation did not cause the loss of the haemagglutinating activity but was sufficient to permit detection of the lectin by streptavidin. A flow cytometric analysis was used for the detection of the receptors on the surface of erythrocytes.
Berger
S
S
Institut de Biotechnologie, Université de Limoges, France.
Karamanos
Y
Y
Schoentgen
F
F
Julien
R
R
D018830:Adhesins, Escherichia coli; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000737:chemistry*; D000954:Antigens, Surface / Q000096:biosynthesis / Q000737:chemistry*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry*; D001427:Bacterial Toxins; D001710:Biotin; D002240:Carbohydrate Sequence; D004912:Erythrocytes / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry*; D005679:G(M3) Ganglioside / Q000031:analogs & derivatives / Q000032:analysis; D006736:Horses; D037102:Lectins / Q000737:chemistry*; D008239:Lysine / Q000032:analysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis
D018830:Adhesins, Escherichia coli; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001427:Bacterial Toxins; D005679:G(M3) Ganglioside; C031429:K99 antigen; D037102:Lectins; D010455:Peptides; C044298:N-glycolylneuraminyllactosylceramide; D001710:Biotin; D008239:Lysine