Evaluation of receptor binding specificity of Escherichia coli K88 (F4) fimbrial adhesin variants using porcine serum transferrin and glycosphingolipids as model receptors.
Diarrheal disease caused by enterotoxigenic Escherichia coli expressing the K88 (F4) fimbrial adhesin (K88 ETEC) is a significant source of mortality and morbidity among newborn and weaned piglets. K88 fimbrial adhesins are filamentous surface appendages whose lectin (carbohydrate-binding) activity allows K88 ETEC to attach to specific glycoconjugates (receptors) on porcine intestinal epithelial cells. There are three variants of K88 adhesin (K88ab, K88ac, and K88ad), which possess different, yet related, carbohydrate-binding specificities. We used porcine serum transferrin (pSTf) and purified glycosphingolipids (GSL) to begin to define the minimal recognition sequence for K88 adhesin variants. We found that K88ab adhesin binds with high affinity to pSTf (dissociation constant, 75 microM), while neither K88ac nor K88ad adhesin recognizes pSTf. Degradation of the N-glycan on pSTf by extensive metaperiodate treatment abolished its interaction with the K88ab adhesin, indicating that the K88ab adhesin binds to the single N-glycan found on pSTf. Using exoglycosidase digestion of the pSTf glycan, we demonstrated that K88ab adhesin recognizes N-acetylglucosamine (GlcNAc) residues in the core of the N-glycan on pSTf. All three K88 variants were found to bind preferentially to GSL containing a beta-linked N-acetylhexosamine (HexNAc), either GlcNAc or N-acetylgalactosamine, in the terminal position or, alternatively, in the penultimate position with galactose in the terminal position. Considering the results from pSTf and GSL binding studies together, we propose that the minimal recognition sequence for the K88 adhesin variants contains a beta-linked HexNAc. In addition, the presence of a terminal galactose beta-linked to this HexNAc residue enhances K88 adhesin binding.
Grange
Philippe A
PA
Veterinary Science Department, South Dakota State University, Brookings, South Dakota 57007, USA.
Mouricout
Michèle A
MA
Levery
Steven B
SB
Francis
David H
DH
Erickson
Alan K
AK
D018830:Adhesins, Escherichia coli / Q000378:metabolism*; D000818:Animals; D006028:Glycosphingolipids / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013552:Swine; D014168:Transferrin / Q000378:metabolism*
D018830:Adhesins, Escherichia coli; D006028:Glycosphingolipids; D014168:Transferrin