The binding of natural and fluorescent lysophospholipids to wild-type and mutant rat liver fatty acid-binding protein and albumin.
Rat liver fatty acid-binding protein (FABP) is able to bind a wide range of non-polar anionic ligands, including lysophospholipids. In order to understand the nature of lysophospholipid interactions with liver FABP, the binding of natural lysophospholipids and two fluorescent analogues, N-(5-dimethylaminonaphthalenesulphonyl)-1-palmitoyl-sn-glycero-3- phosphoethanolamine (dansyl lysoPE) and 1-(O-[11-(5-dimethylaminonaphthalene-sulphonyl)amino]undecyl)-sn-glycero -3- phosphocholine (dansyl-C11-lysoPAF), has been investigated. The results confirmed the ability of liver FABP to bind lysophospholipids with KD values in the range of 1-2 microM, and a 1:1 binding stoichiometry was indicated. Binding of fluorescent lysophospholipids was enhanced with the FABP mutant, R122Q, possibly due to increased flexibility of the binding cavity as a result of reduced hydrogen-bonding constraints. The fluorescent lysophospholipids also bound to albumin, with KD values in the range 0.1-1.0 microM, and could be displaced by oleic acid. The fluorescence characteristics of the dansyl lysophospholipid analogue dansyl-C11-lyso-PAF suggested that this probe binds to the same site(s) on albumin as the fluorescent fatty acid probe 11-(5-dimethylaminonaphthalene-sulphonylamino)-undecanoic acid (DAUDA).
Thumser
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AE
Department of Biochemistry, University of Southampton, U.K.
Wilton
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DC
D000418:Albumins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003619:Dansyl Compounds / Q000378:metabolism; D004735:Energy Transfer; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000378:metabolism; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D005470:Fluorometry; D006860:Hydrogen Bonding; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008246:Lysophospholipids / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014364:Tryptophan / Q000737:chemistry
C093005:1-(O-(11-(5-dimethylaminonaphthalenesulfonyl)amino)undecyl)-sn-glycero-3-phosphocholine; D000418:Albumins; D002352:Carrier Proteins; D003619:Dansyl Compounds; C490174:Fabp7 protein, rat; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D005456:Fluorescent Dyes; D008246:Lysophospholipids; C093004:N-(5-dimethylaminonaphthalenesulfonyl)-1-palmitoyl-sn-glycero-3-phosphoethanolamine; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins; C054400:11-(dansylamino)undecanoic acid; D014364:Tryptophan