A surface-associated retinol- and fatty acid-binding protein (Gp-FAR-1) from the potato cyst nematode Globodera pallida:: lipid binding activities, structural analysis and expression pattern

被引:100
作者
Prior, A [1 ]
Jones, JT
Blok, VC
Beauchamp, J
McDermott, L
Cooper, A
Kennedy, MW
机构
[1] Scottish Crop Res Inst, Mycol & Bacteriol & Nematol Unit, Dundee DD2 5DA, Scotland
[2] Univ Glasgow, Inst Biomed & Life Sci infimm, Div Infect & Immun, Glasgow G12 8QQ, Lanark, Scotland
[3] Univ Glasgow, Dept Chem, Glasgow G12 8QQ, Lanark, Scotland
关键词
host-parasite interaction; lipoxygenase; plant defence; secretions;
D O I
10.1042/0264-6021:3560387
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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-alpha -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 C-11 to C-24, with optimal binding at C-15. 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.
引用
收藏
页码:387 / 394
页数:8
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