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© Thomas Wollert
Publication : Molecular informatics

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

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Molecular informatics - 01 Aug 2013

Savar NS, Sardari S, Jahanian-Najafabadi A, Bouzari S,

Link to Pubmed [PMID] – 27480063

Link to DOI – 10.1002/minf.201300025

Mol Inform 2013 Aug; 32(8): 707-16

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.

https://pubmed.ncbi.nlm.nih.gov/27480063