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Fluorescent dyes as probes to study lipid‐binding proteins
Author(s) -
Pastukhov Alexander V.,
Ropson Ira J.
Publication year - 2003
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.10401
Subject(s) - fluorescence , chemistry , binding affinities , affinities , binding site , dna binding protein , biophysics , plasma protein binding , binding protein , molecule , biochemistry , biology , receptor , transcription factor , organic chemistry , physics , gene , quantum mechanics
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. Proteins 2003;53:000–000. © 2003 Wiley‐Liss, Inc.