The FAR proteins of filarial nematodes: secretion, glycosylation and lipid binding characteristics.
The FAR proteins of nematodes are small ( approximately 20 kDa), helix-rich, fatty acid and retinol-binding (FAR) proteins that appear to be confined to nematodes. We have carried out a comparative sequence and biochemical analysis of selected FAR proteins often species of filarial parasites (from the genera Onchocerca, Brugia, Wuchereria, Loa, Acanthocheilonema and Litomosoides). The sequences fall into two main groups corresponding broadly to the onchocercal and lymphatic filariasis parasites, and only those with unsheathed microfilariae were found to produce glycosylated FAR proteins. The proteins were released into culture medium by all the species and developmental stages investigated. Recombinant forms of two of these proteins (Ov-FAR-1 from O. volvulus and Bm-FAR-1 from B. malayi) were compared for ligand binding in fluorescence-based assays. Both were found to bind all-trans-retinol, (dansylamino) undecanoic acid (DAUDA), and oleic acid by competition. Both produced an identical, and dramatic, blue-shift in the fluorescence emission of DAUDA (from 541 to approximately 483 nm), indicative of similarity in the binding site environments of the two proteins. These findings indicate that there is strong conservation of the biochemical activities of the FAR proteins between the different parasite species, although they appear to have different post-translational modifications which may relate to the biology of the larvae.
Garofalo
Antonio
A
School of Life and Environmental Sciences, University of Nottingham, University Park, UK.
Kläger
Sabine L
SL
Rowlinson
Marie Claire
MC
Nirmalan
Niroshini
N
Klion
Amy
A
Allen
Judith E
JE
Kennedy
Malcolm W
MW
Bradley
Janette E
JE
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary; D005227:Fatty Acids / Q000378:metabolism*; D005370:Filarioidea / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D010802:Phylogeny; D012177:Retinol-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D002352:Carrier Proteins; D018076:DNA, Complementary; D005227:Fatty Acids; D015801:Helminth Proteins; D008024:Ligands; D012177:Retinol-Binding Proteins