title: The cell-binding domain of intimin from enteropathogenic Escherichia coli binds to beta1 integrins.
abstract: Bacteria interact with mammalian cells surface molecules, such as integrins, to colonize tissues and evade immunological detection. Herein, the ability of intimin, an outer membrane protein from enteropathogenic Escherichia coli, to bind beta1 integrins was investigated. Solid-phase binding assays revealed binding of the carboxyl-terminal 280 amino acids of intimin (Int280) to alpha4beta1 and alpha5beta1 integrins. The binding required divalent ions (in particular, it was enhanced by Mn2+) and was inhibited by an RGD-containing peptide. Nonderivatized Int280, but not Int280CS (like Int280 but with Cys-937 replaced by Ser) blocked the binding of biotinylated Int280 to integrins. Int280 did not efficiently inhibit beta1 integrin binding of invasin from Yersinia pseudotuberculosis. Both intimin and invasin, immobilized on plastic surfaces, mediated adherence of resting or phorbol 12-myristate 13-acetate-activated human CD4(+) T cells, whereas fibronectin mediated the adherence of only activated T cells. T cell binding to intimin and invasin was integrin mediated because it was specifically blocked by an RGD-containing peptide and by antibodies directed against the integrin subunits beta1, alpha4, and alpha5. These results demonstrate a specific integrin binding activity for intimin that is related to, but distinct from, that of invasin.
authors.affiliation: Department of Biochemistry, Imperial College of Science, Technology and Medicine, London SW7 2AZ, United Kingdom.
authors.forename: G
authors.initials: G
authors.lastname: Frankel
authors.forename: O
authors.initials: O
authors.lastname: Lider
authors.forename: R
authors.initials: R
authors.lastname: Hershkoviz
authors.forename: A P
authors.initials: AP
authors.lastname: Mould
authors.forename: S G
authors.initials: SG
authors.lastname: Kachalsky
authors.forename: D C
authors.initials: DC
authors.lastname: Candy
authors.forename: L
authors.initials: L
authors.lastname: Cahalon
authors.forename: M J
authors.initials: MJ
authors.lastname: Humphries
authors.forename: G
authors.initials: G
authors.lastname: Dougan
mesh_terms: D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology; D002352:Carrier Proteins; D002448:Cell Adhesion; D004926:Escherichia coli / Q000472:pathogenicity*; D029968:Escherichia coli Proteins; D006801:Humans; D039423:Integrin alpha1; D039441:Integrin alpha4; D039482:Integrin alpha5; D019012:Integrin beta1 / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010455:Peptides / Q000378:metabolism; D011485:Protein Binding
chemical_list: D018829:Adhesins, Bacterial; D015703:Antigens, CD; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D039423:Integrin alpha1; D039482:Integrin alpha5; D019012:Integrin beta1; D009842:Oligopeptides; D010455:Peptides; C053476:invasin, Yersinia; D039441:Integrin alpha4; C073819:eaeA protein, E coli; C047981:arginyl-glycyl-aspartic acid