title: A proteinaceous fraction of wheat bran may interfere in the attachment of enterotoxigenic E. coli K88 (F4+) to porcine epithelial cells.
abstract: Wheat bran (WB) from Triticum aestivum has many beneficial effects on human health. To the best of our knowledge, very little has been published about its ability to prevent pathogenic bacterial adhesion in the intestine. Here, a WB extract was fractionated using different strategies, and the obtained fractions were tested in different in vitro methodologies to evaluate their interference in the attachment of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal porcine epithelial cells (IPEC-J2) with the aim of identifying the putative anti-adhesive molecules. It was found that a proteinaceous compound in the >300-kDa fraction mediates the recognition of ETEC K88 to IPEC-J2. Further fractionation of the >300-kDa sample by size-exclusion chromatography showed several proteins below 90 kDa, suggesting that the target protein belongs to a high-molecular-weight (MW) multi-component protein complex. The identification of some relevant excised bands was performed by mass spectrometry (MS) and mostly revealed the presence of various protease inhibitors (PIs) of low MW: Serpin-Z2B, Class II chitinase, endogenous alpha-amylase/subtilisin inhibitor and alpha-amylase/trypsin inhibitor CM3. Furthermore, an incubation of the WB extract with ETEC K88 allowed for the identification of a 7S storage protein globulin of wheat, Globulin 3 of 66 kDa, which may be one of the most firmly attached WB proteins to ETEC K88 cells. Further studies should be performed to gain an understanding of the molecular recognition of the blocking process that takes place. All gathered information can eventually pave the way for the development of novel anti-adhesion therapeutic agents to prevent bacterial pathogenesis.
authors.affiliation: Servei de Nutrició i Benestar Animal (SNiBA), Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Gemma
authors.initials: G
authors.lastname: González-Ortiz
authors.affiliation: Servei de Proteòmica i Biologia Estructural, Universitat Autònoma de Barcelona, Mòdul B Parc de Recerca, Barcelona, Spain.
authors.forename: Sílvia
authors.initials: S
authors.lastname: Bronsoms
authors.affiliation: Department of Medicinal Chemistry & Chemical Biology, Utrecht University, Utrecht, The Netherlands.
authors.forename: H C
authors.initials: HC
authors.lastname: Quarles Van Ufford
authors.affiliation: Department of Medicinal Chemistry & Chemical Biology, Utrecht University, Utrecht, The Netherlands.
authors.forename: S Bart A
authors.initials: SB
authors.lastname: Halkes
authors.affiliation: Department of Biosciences, General Microbiology, University of Helsinki, Helsinki, Finland.
authors.forename: Ritva
authors.initials: R
authors.lastname: Virkola
authors.affiliation: Department of Medicinal Chemistry & Chemical Biology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Rob M J
authors.initials: RM
authors.lastname: Liskamp
authors.affiliation: Department of Medicinal Chemistry & Chemical Biology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Cees J
authors.initials: CJ
authors.lastname: Beukelman
authors.affiliation: Department of Medicinal Chemistry & Chemical Biology, Utrecht University, Utrecht, The Netherlands.
authors.forename: Roland J
authors.initials: RJ
authors.lastname: Pieters
authors.affiliation: Servei de Nutrició i Benestar Animal (SNiBA), Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: José Francisco
authors.initials: JF
authors.lastname: Pérez
authors.affiliation: Servei de Nutrició i Benestar Animal (SNiBA), Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Barcelona, Spain.
authors.forename: Susana María
authors.initials: SM
authors.lastname: Martín-Orúe
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects*; D002478:Cells, Cultured; D005591:Chemical Fractionation; D004043:Dietary Fiber / Q000494:pharmacology*; D054307:Enterotoxigenic Escherichia coli / Q000502:physiology*; D004847:Epithelial Cells / Q000382:microbiology*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D013552:Swine; D014908:Triticum / Q000737:chemistry
chemical_list: D004043:Dietary Fiber; D010940:Plant Proteins