Parasitic nematodes produce at least two structurally novel classes of small helix-rich retinol- and fatty-acid-binding proteins that have no counterparts in their plant or animal hosts and thus represent potential targets for new nematicides. Here we describe a protein (Gp-FAR-1) from the plant-parasitic nematode Globodera pallida, which is a member of the nematode-specific fatty-acid- and retinol-binding (FAR) family of proteins but localizes to the surface of this species, placing it in a strategic position for interaction with the host. Recombinant Gp-FAR-1 was found to bind retinol, cis-parinaric acid and the fluorophore-tagged lipids 11-(dansylamino)undecanoic acid and dansyl-d,l-α-amino-octanoic acid. The fluorescence emission characteristics of the dansylated analogues indicated that the entire ligand enters the binding cavity. Fluorescence competition experiments showed that Gp-FAR-1 binds fatty acids in the range C11 to C24, with optimal binding at C15. Intrinsic fluorescence analysis of a mutant protein into which a tryptophan residue had been inserted supported computer-based predictions of the position of this residue at the protein's interior and possibly also at the binding site. Of direct relevance to plant defence systems was the observation that Gp-FAR-1 binds two lipids (linolenic and linoleic acids) that are precursors of plant defence compounds and the jasmonic acid signalling pathway. Moreover, Gp-FAR-1 was found to inhibit the lipoxygenase-mediated modification of these substrates in vitro. Thus not only does Gp-FAR-1 function as a broad-spectrum retinol- and fatty-acid-binding protein, the results are consistent with the idea that Gp-FAR-1 is involved in the evasion of primary host plant defence systems.
- host–parasite interaction
- plant defence
↵2 Present address: Department of Nutritional Sciences, Rutgers University, 96 Lipman Drive, New Brunswick, NJ 08901, U.S.A.
The nucleotide sequence data reported will appear in DDBJ, EMBL and GenBank® Nucleotide Sequence Databases under the accession number Y09293.
Abbreviations used: DACA, dansyl-d,l-α-amino-octanoic acid; DAUDA, 11-(dansylamino)undecanoic acid; FAR, fatty-acid- and retinol-binding protein; Gp-FAR-1, FAR from Globodera pallida; J2, second-stage juvenile; rGp-FAR-1, bacterial recombinant Gp-FAR-1 protein.
- The Biochemical Society, London ©2001