Investigating the Role of the Nematode-Specific FAR Family of Proteins

Abstract

Nematode genomes encode for proteins specific to the phylum Nematoda, which bear crucial importance for understanding nematode biology. Despite the importance of nematode-specific proteins and protein families, few have been identified and very little information regarding them is available on public databases. Our work involves a novel, nematode-specific family of proteins, the fatty acid and retinol binding (FAR) proteins of the phylum Nematoda. It was recently discovered that parasitic nematodes of plants, animals and humans secrete these structurally novel FAR proteins into the tissues they occupy. This family of proteins has been shown to be highly conserved across both plant and animal parasitic nematodes, yet there is limited information available regarding this family of proteins and their in vivo function(s) are poorly understood. Our specific research goals are to evaluate the ligand binding properties of FAR protein mutants created via multi site-directed mutagenesis (MSDM). Constructs that do not bind the most commonly screened ligands for FAR proteins will be used to ascertain data on sequence variation and amino acids imperative for ligand binding. To date, we have successfully created forty-eight mutant gene sequences and completed protein expression and purification of fourteen mutants. Mutant proteins were purified via charged nickel affinity chromatography, and all mutants were present in soluble fractions. We expect that our research on this family of proteins may be significant for a broad range of scientists and agriculturalists in understanding plant and animal parasitism, nematode evolution and parasite-control methods.