Fluorescent dyes as probes to study lipid-binding proteins.
We studied the equilibrium binding of two hydrophobic fluorescent dyes, ANS and bisANS, to four members of a family of intracellular lipid-binding proteins: IFABP, CRABP I, CRABP II, and ILBP. The spectral and binding parameters for the probes bound to the proteins were determined. Typically, there was a single binding site on each protein for the ligands. However, IFABP cooperatively bound a second bisANS molecule in the binding pocket. Comparative analysis of affinities and spectral characteristics for the two probes allowed us to examine the contributions of electrostatic and hydrophobic interactions to the binding process, and to address some aspects of the internal structure of the studied proteins.
Department of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Alexander V
AV
Pastukhov
Ira J
IJ
Ropson
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D050556:Fatty Acid-Binding Proteins; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry; D008958:Models, Molecular; D009363:Neoplasm Proteins; D029363:Organic Anion Transporters, Sodium-Dependent; D011485:Protein Binding; D018168:Receptors, Retinoic Acid / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D055672:Static Electricity; D027981:Symporters
D000817:Anilino Naphthalenesulfonates; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D005456:Fluorescent Dyes; D008055:Lipids; D009363:Neoplasm Proteins; D029363:Organic Anion Transporters, Sodium-Dependent; D018168:Receptors, Retinoic Acid; D027981:Symporters; C082927:retinoic acid binding protein I, cellular; C082928:retinoic acid binding protein II, cellular; C071908:sodium-bile acid cotransporter; C027132:1-anilino-8-naphthalenesulfonate; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate)