Innate immune signaling inhibition via disruption of adapter interactions.

Abstract

Abstract Toll-like receptors (TLR) form homo- or heterodimers in response to pathogen-associated molecular patterns. The TLR homo- or heterodimers establish an intracellular signaling platform that recruits the signaling adapter protein Toll-Interleukin 1 Receptor (TIR) domain containing adaptor protein (TIRAP). This receptor-mediated platform initiates a signaling cascade which leads to NF-kB activation, cytokine production, and inflammation. Inflammation leads to pathogen clearance; however, excessive stimulation of the innate immune pathway can lead to injury and death. We are investigating mechanisms to disrupt TIRAP-mediated TLR signaling via two mechanisms: the bacterial TIR domain protein, TcpB from Brucella sp., and the decoy peptide, 2R9. We have previously determined the structures of TIRAP and TcpB. Presumably via molecular interactions with TIRAP, TcpB inhibited NF-kB activation. We now report surface plasma resonance binding of TIRAP with TcpB. The decoy peptide, 2R9, was derived from the TLR2 TIR domain D-helix. We have previously described interactions between 2R9 and TIRAP via surface plasma resonance. Based on the ability of 2R9 peptide to protect against lethality in response to acute influenza infection, 2R9 peptide could highlight a novel drug target to prevent injury and death caused by innate immune over activation. Using recombinant TIRAP with TcpB and 2R9 peptide, we are performing structural studies to characterize molecular interactions to identify putative sites for therapeutic intervention.