title: In Silico and In Vitro Studies of Truncated Forms of Flagellin (FliC) of Enteroaggregative Escherichia coli (EAEC).
abstract: Enteroaggregative Escherichia coli (EAEC) is an important cause of acute and chronic diarrhea worldwide. It has been shown that flagellin (FliC), a major bacterial surface protein of EAEC, causes IL-8 release from certain epithelial cell lines via activation of TLR-5. Based on the ability of this protein to activate innate immunity, flagellin can be considered as a potent adjuvant in new vaccines and adjuvant effects of native or recombinant forms of flagellin have been demonstrated. In the current study we designed various truncated forms of FliC-EAEC based on its interaction site with TLR-5 and assessed the interactions via docking protocols. Then, the most appropriate truncated forms were PCR amplified, cloned in to pGEX-5X-1 plasmid and expressed. Finally, the expressed proteins were tested for pro-inflammatory properties. Our in silico and in vitro results indicated that two truncated forms of FliC- EAEC (amino acids 79-117 and 477-508) effectively interact with TLR-5, thus could be capable of inducing in vivo immune responses.
authors.affiliation: Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619.
authors.forename: Nastaran Sadat
authors.initials: NS
authors.lastname: Savar
authors.affiliation: Bioinformatics and Drug Design Unit, Department of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran, 13164, Tel: (+98-21) 6648-0780; Fax: (+98-21) 6648-0780. ssardari@hotmail.com.
authors.forename: Soroush
authors.initials: S
authors.lastname: Sardari
authors.affiliation: Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619.|Present address: Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Jahanian-Najafabadi
authors.affiliation: Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619. saeidbouzari@yahoo.com.
authors.forename: Saeid
authors.initials: S
authors.lastname: Bouzari
keywords: Enteroaggregative Escherichia coli; FliC; In silico study; Truncated forms