Biochemical and biophysical characterization of Dr-DLP-1, the major pseudocoelomic lipid binding protein of the giant kidney worm Dioctophyme renale.

Adults of the parasitic nematode Dioctophyme renale locate in the renal pelvis of one (usually the right) kidney of mammals, destroying its parenchyma and function. The unaffected kidney compensates and hypertrophies such that, in most cases, there are no clear clinical signs of infection. It has recently been shown that the pseudocelomic body fluid of D. renale contains a highly abundant protein of 44 kDa in both males and females, here dubbed "dorylipophorin" (Dr-DLP-1), that binds lipids in highly apolar sites. Orthologues of this protein are specific to Clade I (Dorylaimia) of the Phylum Nematoda, initially described as the poly-cysteine and histidine-tailed proteins of unknown function of Trichinella spiralis, and one that is the immunomodulatory secreted p43 protein from Trichuris muris. We here present a biochemical and biophysical characterization of Dr-DLP-1, demonstrating that it is N-glycosylated, is more stable when bound to a fatty acid, and can be detected in the fluid surrounding the parasite in parasitized kidneys. The analysis of Dr-DLP-1 lipid binding activity showed Kd values of 2.1 ± 0.2 μM and 2.2 ± 0.4 μM for the fluorescent probes 11-(dansylamino) undecanoic acid (DAUDA) and 8-anilinonaphthalene-1-sulfonic acid (ANS) respectively. Moreover, the apparent affinity constants for oleic acid (Kdapp 6.9 ± 0.5 μM) and cholesterol (Kdapp 27.6 ± 2 μM) were estimated by competition assays. As a whole, these results indicate that Dr-DLP-1 binds fatty acids with an affinity appropriate for a transporter. This novel nematode protein likely has a role in mass distribution of lipids within the parasites and could be functionally replacing the nematode polyprotein allergens (NPAs) found in other nematode clades.