@article{mbs:/content/journal/micro/10.1099/00221287-148-3-657, author = "Reece, Stephen and Simmons, Cameron P and Fitzhenry, Robert J and Batchelor, Miranda and Hale, Christine and Matthews, Stephen and Phillips, Alan D and Dougan, Gordon and Frankel, Gad", title = "Mutagenesis of conserved tryptophan residues within the receptor-binding domain of intimin: influence on binding activity and virulence", journal= "Microbiology", year = "2002", volume = "148", number = "3", pages = "657-665", doi = "https://doi.org/10.1099/00221287-148-3-657", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-148-3-657", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "EPEC, EHEC, Tir", keywords = "LEE, locus of enterocyte effacement", keywords = "Citrobacter rodentium", keywords = "EPEC, enteropathogenic Escherichia coli", keywords = "A/E lesion, ‘attaching and effacing’ lesion", keywords = "EHEC, enterohaemorrhagic Escherichia coli", keywords = "MBP, maltose-binding protein", keywords = "FAS, fluorescent actin stain", keywords = "Tir, translocated intimin receptor", abstract = "Intimate bacterial adhesion to intestinal epithelium is a pathogenic mechanism shared by several human and animal enteric pathogens, including enteropathogenic and enterohaemorrhagic Escherichia coli and Citrobacter rodentium. The proteins directly involved in this process are the outer-membrane adhesion molecule intimin and the translocated intimin receptor, Tir. The receptor-binding activity of intimin resides within the carboxy terminus 280 aa (Int280) of the polypeptide. Four tryptophan residues, W117/776, W136/795, W222/881 and W240/899, are conserved within different Int280 molecules that otherwise show considerable sequence variation. In this study the influence of site-directed mutagenesis of each of the four tryptophan residues on intimin-Tir interactions and on intimin-mediated intimate attachment was determined. The mutant intimins were also studied using a variety of in vitro and in vivo infection models. The results show that all the substitutions modulated intimin activity, although some mutations had more profound effects than others.", }