Studies on fatty-acid-binding proteins. The purification of rat liver fatty-acid-binding protein and the role of cysteine-69 in fatty acid binding.
1. A new, simple and high-yield procedure is described for the purification of hepatic fatty-acid-binding protein from rat liver using naphthylaminodecyl-agarose as an affinity column. 2. Cysteine-69 is shown to react slowly, but quantitatively, with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), indicating that the thiol group is free, but may be buried within the protein. 3. Fatty acids do not affect the DTNB reactivity of this cysteine residue; however, cysteine reactivity is enhanced in the presence of haem and oleoyl-CoA. 4. Fatty-acid-binding protein that has been modified with DTNB is still able to bind the fluorescent fatty acid 11-(dansylamino)undecanoic acid, indicating that cysteine-69 may be remote from the fatty-acid-binding site.
Wilton
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DC
Department of Biochemistry, School of Biochemical and Physiological Sciences, University of Southampton, U.K.
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003545:Cysteine / Q000502:physiology*; D004228:Dithionitrobenzoic Acid; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000378:metabolism*; D006418:Heme / Q000378:metabolism; D008024:Ligands; D008099:Liver / Q000032:analysis*; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D051381:Rats; D011919:Rats, Inbred Strains
D002352:Carrier Proteins; 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; D006418:Heme; D004228:Dithionitrobenzoic Acid; D003545:Cysteine