Lightning Talk (10 min oral) The 48th Lorne Conference on Protein Structure and Function 2023

Anti-Inflammatory Strategies Through Engineering of Natural Chemokine-Binding Proteins (#28)

Ram Bhusal 1 , Shankar Devkota 1 , Pramod Aryal 1 , Richard J Payne 2 , Matthew CJ Wilce 1 , Martin J Stone 1
  1. Infection and Immunity Program, Monash Biomedicine Discovery Institute, and Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia
  2. School of Chemistry, The University of Sydney, Sydney, NSW, Australia

A hallmark of chronic inflammatory diseases is the excessive accumulation of white blood cells in the affected tissues, which is coordinated by pro-inflammatory mediators called chemokines. Humans have ~50 chemokines divided into two major classes – CC and CXC chemokines. Specific groups of chemokines are associated with different inflammatory diseases; for example, the chemokines CCL2, CCL7 and CCL8 are involved in atherosclerosis. As natural chemokine inhibitors, evasin proteins secreted in tick saliva are potential anti-inflammatory therapeutic agents. However, the development of tick evasins as chemokine-targeted anti-inflammatory therapeutics requires an understanding of the factors controlling their chemokine-binding specificity. Structures of the evasins EVA-P974 and EVA-AAM02 bound to several human CC chemokine ligands (CCL7, CCL11, CCL16 and CCL17) and to a CCL8-CCL7 chimera reveal that the specificity of evasins for chemokines of the CC subfamily is defined by conserved, rigid backbone-backbone interactions.  Whereas the preference for a subset of CC chemokines is controlled by side chain interactions at hotspots in flexible structural elements. The substitution of amino acid residues at hotspots provides evasin variants with new chemokine-binding properties. The structures of engineered evasins bound to several chemokines reveal an underlying molecular mechanism. This molecular understanding enabled us to rationally engineer evasins with modified chemokine selectivity, providing a foundation for the future engineering of evasins as anti-inflammatory therapeutics.

References.

  1. Bhusal, R.P.*; Aryal, P.; Devkota, S.; Pokhrel, R.; Gunzburg, M.J.; Foster, S.R.; Lim, H.D.; Payne, R.J.; Wilce, M.C.J.; Stone, M.J.* Structure-guided engineering of tick evasins for targeting chemokines in inflammatory diseases. Proc. Natl. Acad. Sci. USA 2022, 119. (*corresponding author)
  2. Aryal, P.; Devkota, S.; Jeevarajah, D.; law, R.; Payne, R.; Bhusal, R.P.*; Stone, M.J.* Swapping N-terminal regions among tick evasins reveals cooperative interactions influencing chemokine binding and selectivity. J. Biol. Chem. 2022, 298. (*corresponding author)
  3. Devkota, S.; Aryal, P.; Wilce, M.C.J.; Payne, R.J.; Bhusal, R.P.*; Stone, M.J.* Structural basis of broad-spectrum chemokine inhibition by EVA-AAM02 (Manuscript in Preparation) (*corresponding author)
  4. Bhusal, R.P.; Eaton, J.; Chowdhury, S.T.; Power, C.A.; Proudfoot, A.E.I.; Bhattacharya, S.; Stone, M. J. Evasins: tick salivary proteins that inhibit mammalian chemokines. TrendsBiochem. Sci. 2020, 45, 108.