A Proteinaceous Fraction of Wheat Bran May Interfere in the Attachment of Enterotoxigenic E. Coli K88 (F4+) to Porcine Epithelial Cells
González-Ortiz, Gemma (Universitat Autònoma de Barcelona. Servei de Nutrició i Benestar Animal)
Bronsoms, Sílvia (Universitat Autònoma de Barcelona. Servei de Proteòmica i Biologia Estructural)
Quarles Van Ufford, H. C. (Utrecht University. Department of Medicinal Chemistry & Chemical Biology)
Halkes, S. Bart A. (Utrecht University. Department of Medicinal Chemistry & Chemical Biology)
Virkola, Ritva (University of Helsinki. Department of Biosciences, General Microbiology)
Liskamp, Rob M. J. (Utrecht University. Department of Medicinal Chemistry & Chemical Biology)
Beukelman, Cees J. (Utrecht University. Department of Medicinal Chemistry & Chemical Biology)
Pieters, Roland J. (Utrecht University. Department of Medicinal Chemistry & Chemical Biology)
Pérez Hernández, José Francisco (Universitat Autònoma de Barcelona. Servei de Nutrició i Benestar Animal)
Martín Orúe, Susana M. (Universitat Autònoma de Barcelona. Servei de Nutrició i Benestar Animal)

Data:	2014
Resum:	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.
Ajuts:	Ministerio de Educación y Ciencia AGL2009-07328
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Pili and fimbriae ; Protein extraction ; Adhesives ; Common Wheat ; Wheat ; Globulins ; Bacterial Pathogens ; Carbohydrates
Publicat a:	PloS one, Vol. 9 (august 2014) , ISSN 1932-6203

DOI: 10.1371/journal.pone.0104258
PMID: 25119298

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