title: N-terminal truncation enables crystallization of the receptor-binding domain of the FedF bacterial adhesin.
abstract: FedF is the two-domain tip adhesin of F18 fimbriae from enterotoxigenic Escherichia coli. Bacterial adherence, mediated by the N-terminal receptor-binding domain of FedF to carbohydrate receptors on intestinal microvilli, causes diarrhoea and oedema disease in newly weaned piglets and induces the secretion of Shiga toxins. A truncate containing only the receptor-binding domain of FedF was found to be further cleaved at its N-terminus. Reconstruction of this N-terminal truncate rendered FedF amenable to crystallization, resulting in crystals with space group P2(1)2(1)2(1) and unit-cell parameters a = 36.20, b = 74.64, c = 99.03 A that diffracted to beyond 2 A resolution. The binding specificity of FedF was screened for on a glycan array, exposing 264 glycoconjugates, to identify specific receptors for cocrystallization with FedF.
authors.affiliation: Department of Ultrastructure, Vrije Universiteit Brussel, Flanders Interuniversity Institute for Biotechnology, Pleinlaan 2, 1050 Brussels, Belgium.
authors.forename: Maia
authors.initials: M
authors.lastname: De Kerpel
authors.forename: Inge
authors.initials: I
authors.lastname: Van Molle
authors.forename: Lea
authors.initials: L
authors.lastname: Brys
authors.forename: Lode
authors.initials: L
authors.lastname: Wyns
authors.forename: Henri
authors.initials: H
authors.lastname: De Greve
authors.forename: Julie
authors.initials: J
authors.lastname: Bouckaert
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011971:Receptors, Immunologic / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D018829:Adhesins, Bacterial; D029968:Escherichia coli Proteins; C440653:FedF protein, E coli; D010446:Peptide Fragments; D011971:Receptors, Immunologic; C047105:bacterial adhesin receptor