Localization of lysine residues in the binding domain of the K99 fibrillar subunit of enterotoxigenic Escherichia coli.
Modification of lysine residues with 4-chloro-3,5-dinitrobenzoate results in the loss of the binding capacity of K99 fibrillae to horse erythrocytes (Jacobs, A.A.C., van Mechelen, J.R. and de Graaf, F.K. (1985) Biochim. Biophys. Acta 832, 148-155). In the present study we used dinitrobenzoate as a spectral probe to map the modified residues. After the incorporation of 0.7 mol CDNB per mol subunit, 90% of the binding activity disappeared and the lysine residues at positions 87, 132 and 133 incorporated 20%, 27.5% and 52.2% of the totally incorporated label, respectively. In the presence of the glycolipid receptor, Lys-132 and Lys-133 were partially protected against modification, while Lys-87 was not protected. The results suggest that Lys-132 and Lys-133 are part of the receptor-binding domain of the K99 fibrillar subunit and that the positive charges on these residues are important for the interaction of the fibrillae with the negatively charged sialic acid residue of the glycolipid receptor. A striking homology was found between a six-amino-acid residue segment of K99, containing Lys-132 and Lys-133, and segments of three other sialic-acid-specific lectins; cholera toxin B subunit, heat-labile toxin B subunit of Escherichia coli and CFA1 fimbrial subunit, suggesting that these segments might also be part of the receptor-binding domain in these three proteins.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001665:Binding Sites; D002723:Chlorobenzoates / Q000494:pharmacology; D002918:Chymotrypsin / Q000378:metabolism; D004912:Erythrocytes / Q000382:microbiology; D004926:Escherichia coli / Q000378:metabolism*; D006736:Horses; D008239:Lysine / Q000032:analysis*; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000032:analysis
D000596:Amino Acids; D001426:Bacterial Proteins; D002723:Chlorobenzoates; D046911:Macromolecular Substances; D010446:Peptide Fragments; C015746:4-chloro-3,5-dinitrobenzoate; D002918:Chymotrypsin; D008239:Lysine