A native 13-kDa fatty acid binding protein from the liver fluke Fasciola hepatica.
A 13-kDa fatty acid binding protein (FABP) (Fh13) has been isolated from the cytosol of adult Fasciola hepatica and its physicochemical and binding characteristics determined. Fh13 appears to exist as a dimer in native solution. Binding of the fluorescent fatty acid analogue 11-((5-dimethyl aminonaphthalene-1-sulfonyl) amino) undecanoic acid (DAUDA) to Fh13 results in changes in the emission spectrum, which are reversed by oleic acid. The binding activity for DAUDA determined from titration experiments revealed a single binding site per monomeric unit with Kd of 1.5 microM. The displacement of DAUDA by competitive nonfluorescent ligands allowed Kd values for oleic (2.5 microM), retinoic (2.8 microM), palmitic (4.1 microM) and arachidonic acid (6.1 microM) to be calculated. Ten commonly used anthelmintics were evaluated for binding to Fh13, but only bithionol showed binding activity commensurate with those of the putative natural ligands (Kd 6.8 microM).
Timanova-Atanasova
Anna
A
Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria.
Jordanova
Rosita
R
Radoslavov
Georgi
G
Deevska
Gergana
G
Bankov
Ilia
I
Barrett
John
J
D000818:Animals; D000871:Anthelmintics / Q000378:metabolism; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D003619:Dansyl Compounds / Q000378:metabolism*; D005210:Fasciola hepatica / Q000737:chemistry* / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D008024:Ligands; D008970:Molecular Weight; D011485:Protein Binding
D000871:Anthelmintics; D002352:Carrier Proteins; D003619:Dansyl Compounds; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D005456:Fluorescent Dyes; D008024:Ligands; C054400:11-(dansylamino)undecanoic acid