Characterization of ligand binding to acyl-CoA-binding protein.
Ligand binding to recombinant bovine acyl-CoA-binding protein (rACBP) was examined using a Lipidex 1000 competition assay and an e.p.r. spectroscopy displacement assay. Of all putative ligands tested, rACBP exhibited a high binding affinity only for acyl-CoA esters. No alternative ligands could be found in rat liver fractions purified on an affinity of column on which ACBP was coupled to Sepharose 4B. E.p.r. data indicate that both the acyl chain and the CoA head group of acyl-CoA are involved in binding and that the 3'-phosphate group on the ribose moiety of acyl-CoA esters plays a crucial role in the binding of acyl-CoA to ACBP. E.p.r. competition binding studies show that the binding affinity of acyl-CoA esters for rACBP is strongly dependent on the length of the acyl chain with a clear preference for acyl-CoA esters with 14-22 carbon atoms in the acyl chain. No correlation between the number of double bonds in the acyl chain and the binding affinity was observed. The experimental results strongly indicate that ACBP specifically binds long-chain acyl-CoA esters with a very high affinity, results that indicate that ACBP is likely to be involved in the intracellular transport and pool formation of these compounds.
Rosendal
J
J
Institute of Biochemistry, Odense University, Denmark.
Ertbjerg
P
P
Knudsen
J
J
D000214:Acyl Coenzyme A / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism*; D002417:Cattle; D003911:Dextrans / Q000378:metabolism; D004952:Esters; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000378:metabolism; D008024:Ligands; D008297:Male; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism
D000214:Acyl Coenzyme A; D002352:Carrier Proteins; D003911:Dextrans; D004952:Esters; C490174:Fabp7 protein, rat; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D008024:Ligands; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; C010599:lipidex