CGC Bibliography Paper 5754

The FAR protein family of the nematode Caenorhabditis elegans - differential lipid binding properties, structural characteristics, and developmental regulation.

Garofalo A, Rowlinson MC, Amambua NA, Hughes JM, Kelly SM, Price NC, Cooper A, Watson DG, Kennedy MW, Bradley JE

Medline:

12502713

Citation:

Journal of Biological Chemistry 278: 8065-8074 2003

Type:

ARTICLE

Genes:

far-1 far-2 far-3 far-4 far-5 far-6 far-7 far-8

Abstract:

Parasitic nematodes of humans and plants secrete a structurally novel type of fatty acid- and retinol-binding protein, FAR, into the tissues they occupy. These proteins may interfere with intercellular lipid signaling to manipulate the defense reactions of the host or acquire essential lipids for the parasites. The genome of the nematode Caenorhabditis elegans encodes eight FAR-like proteins (Ce-FAR-1 to -8). These fall into three discrete groups as indicated by phylogenetic sequence comparisons and intron positions, the proteins from parasitic nematodes falling into group A. Recombinant Ce-FAR-1 to -7 were produced in Escherichia coli and tested for lipid binding in fluorescence-based assays. Ce-FAR-1 to -6 bound DAUDA (11-((5-dimethylaminonaphthalene-1-sulfonyl)amino)undecanoi c acid), cis-parinaric acid, and retinol with dissociation constants in the micromolar range, whereas Ce-FAR-7 bound the latter two lipids relatively poorly. Each protein produced a characteristic shift in peak fluorescence emission of DAUDA, and one (Ce-FAR-5) produced a shift greater than has been observed previously for any lipid-binding protein. Selected Ce-FAR proteins were analyzed by circular dichroism (CD) and differential scanning calorimetry, were, found to be helix-rich, and exhibited high thermal stability (transition midpoint, 82.7 degreesC). CD and secondary structure predictions, however, both indicated that Ce-FAR-7 possesses substantially less helix than the other FAR proteins. The genes encoding the Ce-FAR proteins were found to be transcribed differentially through the life cycle of C. elegans, such that Ce-far-4 was transcribed at highest levels in the fourth larval stage, and Ce-far-3 and -7 predominated in males.