Chemokines are a group of chemotaxis proteins that regulate cell trafficking and play important roles in immune responses and inflammation. Ticks are blood-sucking parasites that secrete numerous immune-modulatory agents in their saliva to evade host immune responses. Evasin-3 is a small salivary protein which belongs to a class of chemokine-binding proteins isolated from the brown dog tick， Rhipicephalus sanguineus. Evasin-3 has been shown to have a high affinity for chemokines CXCL1 and CXCL8 and diminish inflammation in mice. In the present study， solution NMR spectroscopy was used to investigate the structure of Evasin-3 and its CXCL8/Evasin-3 complex. Evasin-3 is found to disrupt the glycosaminoglycan ... More
Chemokines are a group of chemotaxis proteins that regulate cell trafficking and play important roles in immune responses and inflammation. Ticks are blood-sucking parasites that secrete numerous immune-modulatory agents in their saliva to evade host immune responses. Evasin-3 is a small salivary protein which belongs to a class of chemokine-binding proteins isolated from the brown dog tick， Rhipicephalus sanguineus. Evasin-3 has been shown to have a high affinity for chemokines CXCL1 and CXCL8 and diminish inflammation in mice. In the present study， solution NMR spectroscopy was used to investigate the structure of Evasin-3 and its CXCL8/Evasin-3 complex. Evasin-3 is found to disrupt the glycosaminoglycan (GAG) binding site of CXCL8 and inhibit the interaction of CXCL8 with CXCR2. Structural data were used to design two novel CXCL8 binding peptides. Linear tEv3 17-56 and cyclic tcEv3 16-56 dPG Evasin-3 variants were chemically synthesized by solid phase peptide synthesis. Affinity of these newly synthesized variants to CXCL8 were measured by surface plasmon resonance (SPR) biosensor analysis. Kd values of tEv3 17-56 and tcEv3 16-56 dPG were 27 and 13 nM， respectively. Both compounds effectively inhibited CXCL8 induced migration of polymorphonuclear neutrophils (PMN). The present results suggest utility of synthetic Evasin-3 variants as scaffolds for designing and fine-tuning new chemokine-binding agents that suppress immune responses and inflammation.
