Rahman, Hossinur M. King, Rebecca K. Shewell, Lucy A. Semchenko, Evgeny E. Hartley-Tassell, Lauren Wilson, Jennifer C. J. Day, Christopher Korolik, Victoria Characterisation of a Multi-ligand Binding Chemoreceptor CcmL (Tlp3) of <i>Campylobacter jejuni</i> <div><p><i>Campylobacter jejuni</i> is the leading cause of human gastroenteritis worldwide with over 500 million cases annually. Chemotaxis and motility have been identified as important virulence factors associated with <i>C. jejuni</i> colonisation. Group A transducer-like proteins (Tlps) are responsible for sensing the external environment for bacterial movement to or away from a chemical gradient or stimulus. In this study, we have demonstrated Cj1564 (Tlp3) to be a multi-ligand binding chemoreceptor and report direct evidence supporting the involvement of Cj1564 (Tlp3) in the chemotaxis signalling pathway via small molecule arrays, surface plasmon and nuclear magnetic resonance (SPR and NMR) as well as chemotaxis assays of wild type and isogenic mutant strains. A modified nutrient depleted chemotaxis assay was further used to determine positive or negative chemotaxis with specific ligands. Here we demonstrate the ability of Cj1564 to interact with the chemoattractants isoleucine, purine, malic acid and fumaric acid and chemorepellents lysine, glucosamine, succinic acid, arginine and thiamine. An isogenic mutant of <i>cj1564</i> was shown to have altered phenotypic characteristics of <i>C. jejuni</i>, including loss of curvature in bacterial cell shape, reduced chemotactic motility and an increase in both autoagglutination and biofilm formation. We demonstrate Cj1564 to have a role in invasion as in <i>in vitro</i> assays the <i>tlp3</i> isogenic mutant has a reduced ability to adhere and invade a cultured epithelial cell line; interestingly however, colonisation ability of avian caeca appears to be unaltered. Additionally, protein-protein interaction studies revealed signal transduction initiation through the scaffolding proteins CheV and CheW in the chemotaxis sensory pathway. This is the first report characterising Cj1564 as a multi-ligand receptor for <i>C. jejuni</i>, we therefore, propose to name this receptor CcmL, <i><u>C</u>ampylobacter</i><u>c</u>hemoreceptor for <u>m</u>ultiple <u>l</u>igands. In conclusion, this study identifies a novel multifunctional role for the <i>C. jejuni</i> CcmL chemoreceptor and illustrates its involvement in the chemotaxis pathway and subsequent survival of this organism in the host.</p></div> characterisation;multi-ligand;binding;chemoreceptor;ccml 2014-01-02
    https://plos.figshare.com/articles/dataset/Characterisation_of_a_Multi_ligand_Binding_Chemoreceptor_CcmL_Tlp3_of_Campylobacter_jejuni_/892436
10.1371/journal.ppat.1003822