The bundlin pilin protein of enteropathogenic Escherichia coli is an N-acetyllactosamine-specific lectin.
Synthetic N-acetyllactosamine (LacNAc) glycoside sequences coupled to BSA competitively inhibit enteropathogenic Escherichia coli (EPEC) localized adherence (LA) to human intestinal biopsy specimens and tissue culture cell monolayers. The LacNAc-specific adhesin appears to be associated with the bundle-forming pili (BFP) expressed by EPEC during the early stages of colonization. Herein, we report that recombinant bundlin inhibits EPEC LA to HEp-2 cells and binds to HEp-2 cells. Recombinant bundlin also binds, with millimolar association constants (K(assoc)), to synthetic LacNAc-Benzene and LacNAc-O(CH(2))(8)CONH(2) glycosides as assessed in the gas phase by nanoelectrospray ionization mass spectrometry. Furthermore, LacNAc-BSA inhibits LA only of EPEC strains that express alpha bundlin alleles, suggesting putative locations for the LacNAc-binding pocket in the alpha bundlin monomer. Collectively, these results suggest that alpha bundlin possesses lectin-like properties that are responsible for LacNAc-specific initial adherence of alpha bundlin-expressing EPEC strains to host intestinal epithelial cells.
Hyland
Romney M
RM
University of Calgary, Calgary, AB, Canada.
Sun
Jiangxiao
J
Griener
Thomas P
TP
Mulvey
George L
GL
Klassen
John S
JS
Donnenberg
Michael S
MS
Armstrong
Glen D
GD
D000606:Amino Sugars / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000502:physiology*; D002460:Cell Line; D054308:Enteropathogenic Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
D000606:Amino Sugars; D029968:Escherichia coli Proteins; D037102:Lectins; D011994:Recombinant Proteins; D036781:Fimbriae Proteins; C000458:N-acetyllactosamine
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