title: High relaxivity supramolecular adducts between human-liver fatty-acid-binding protein and amphiphilic Gd(III) complexes: structural basis for the design of intracellular targeting MRI probes.
abstract: Gadolinium complexes linked to an apolar fragment are known to be efficiently internalized into various cell types, including hepatocytes. Two lipid-functionalized gadolinium chelates have been investigated for the targeting of the human liver fatty acid binding protein (hL-FABP) as a means of increasing the sensitivity and specificity of intracellular-directed MRI probes. hL-FABP, the most abundant cytosolic lipid binding protein in hepatocytes, displays the ability to interact with multiple ligands involved in lipid signaling and is believed to be an obligate carrier to escort lipidic drugs across the cell. The interaction modes of a fatty acid and a bile acid based gadolinium complex with hL-FABP have been characterized by relaxometric and NMR experiments in solution with close-to-physiological protein concentrations. We have introduced the analysis of paramagnetic-induced protein NMR signal intensity changes as a quantitative tool for the determination of binding stoichiometry and of precise metal-ion-center positioning in protein-ligand supramolecular adducts. A few additional NMR-derived restraints were then sufficient to locate the ligand molecules in the protein binding sites by using a rapid data-driven docking method. Relaxometric and (13)C NMR competition experiments with oleate and the gadolinium complexes revealed the formation of heterotypic adducts, which indicates that the amphiphilic compounds may co-exist in the protein cavity with physiological ligands. The differences in adduct formation between fatty acid and bile acid based complexes provide the basis for an improved molecular design of intracellular targeted probes.
authors: Department of Biotechnology, University of Verona, Strada Le Grazie, 15, 37134 Verona, Italy.
authors: Mariapina
authors: M
authors: D'Onofrio
authors: Eliana
authors: E
authors: Gianolio
authors: Alberto
authors: A
authors: Ceccon
authors: Francesca
authors: F
authors: Arena
authors: Serena
authors: S
authors: Zanzoni
authors: David
authors: D
authors: Fushman
authors: Silvio
authors: S
authors: Aime
authors: Henriette
authors: H
authors: Molinari
authors: Michael
authors: M
authors: Assfalg
mesh_terms: D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002614:Chelating Agents / Q000737:chemistry*; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry* / Q000378:metabolism; D005682:Gadolinium / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008099:Liver / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D015394:Molecular Structure; D011485:Protein Binding; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D002352:Carrier Proteins; D002614:Chelating Agents; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D008055:Lipids; D008562:Membrane Glycoproteins; D013501:Surface-Active Agents; C037643:bile acid binding proteins; D005682:Gadolinium