title: The molecular basis for recognition of bacterial ligands at equine TLR2, TLR1 and TLR6.
abstract: TLR2 recognises bacterial lipopeptides and lipoteichoic acid, and forms heterodimers with TLR1 or TLR6. TLR2 is relatively well characterised in mice and humans, with published crystal structures of human TLR2/1/Pam3CSK4 and murine TLR2/6/Pam2CSK4. Equine TLR4 is activated by a different panel of ligands to human and murine TLR4, but less is known about species differences at TLR2. We therefore cloned equine TLR2, TLR1 and TLR6, which showed over 80% sequence identity with these receptors from other mammals, and performed a structure-function analysis. TLR2/1 and TLR2/6 from both horses and humans dose-dependently responded to lipoteichoic acid from Staphylococcus aureus, with no significant species difference in EC50 at either receptor pair. The EC50 of Pam2CSK4 was the same for equine and human TLR2/6, indicating amino acid differences between the two species' TLRs do not significantly affect ligand recognition. Species differences were seen between the responses to Pam2CSK4 and Pam3CSK4 at TLR2/1. Human TLR2/1, as expected, responded to Pam3CSK4 with greater potency and efficacy than Pam2CSK4. At equine TLR2/1, however, Pam3CSK4 was less potent than Pam2CSK4, with both ligands having similar efficacies. Molecular modelling indicates that the majority of non-conserved ligand-interacting residues are at the periphery of the TLR2 binding pocket and in the ligand peptide-interacting regions, which may cause subtle effects on ligand positioning. These results suggest that there are potentially important species differences in recognition of lipopeptides by TLR2/1, which may affect how the horse deals with bacterial infections.
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB30ES, UK. ceb27@cam.ac.uk.
authors.forename: Katherine Lucy
authors.initials: KL
authors.lastname: Irvine
authors.forename: Lee Jason
authors.initials: LJ
authors.lastname: Hopkins
authors.forename: Monique
authors.initials: M
authors.lastname: Gangloff
authors.forename: Clare Elizabeth
authors.initials: CE
authors.lastname: Bryant
mesh_terms: D000818:Animals; D057809:HEK293 Cells; D006736:Horses / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008024:Ligands; D055666:Lipopeptides / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D051379:Mice; D008968:Molecular Conformation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013211:Staphylococcus aureus / Q000502:physiology; D013682:Teichoic Acids / Q000378:metabolism; D051194:Toll-Like Receptor 1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051216:Toll-Like Receptor 6 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D008024:Ligands; D055666:Lipopeptides; D008070:Lipopolysaccharides; D013682:Teichoic Acids; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2; D051216:Toll-Like Receptor 6; C009900:lipoteichoic acid