In Silico and In Vitro Studies of Truncated Forms of Flagellin (FliC) of Enteroaggregative Escherichia coli (EAEC)

Authors

  • Nastaran Sadat Savar,

    1. Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619
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  • Soroush Sardari,

    Corresponding author
    1. 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
    • 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
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  • Ali Jahanian-Najafabadi,

    1. Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619
    2. Present address: Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran
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  • Saeid Bouzari

    Corresponding author
    1. Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619
    • Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran, 1316943551 tel: (+98-21) 66953311, (ext 2221); fax: (+98-21) 6649-2619
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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.

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