Aromatic side-chain cluster of biotin binding site of avidin allows circular dichroism spectroscopic investigation of its ligand binding properties.
Promiscuous ligand binding by hen egg-white avidin has been demonstrated and studied by using circular dichroism (CD) spectroscopy complemented by molecular docking calculations. It has been shown that the biotin-binding pocket of avidin is able to accommodate a wide variety of chemical compounds including therapeutic drugs (e.g., thalidomide, NSAIDs, antihistamines), natural compounds (bilirubin, myristic acid), and synthetic agents (xanthenone dyes). The cluster of aromatic residues located at the biotin-binding pocket renders the intrinsic CD spectrum of avidin sensitive to ligand binding that results in the increase of the vibronic components of the (1) L(b) transition of the Trp residues. Extrinsic (induced) CD bands measured with chemically diverse avidin ligands are generated by intramolecular coupled oscillator (e.g., bilirubin) or by intermolecular ligand-Trp exciton coupling mechanism [e.g., 2-(4'-hydroxyazobenzene)-benzoic acid (HABA)]. Among the compounds of which avidin-binding affinity constants have been calculated, two novel high-affinity ligands, flufenamic acid and an enzyme inhibitor thiazole derivative have been identified (K(d) ≈ 1 μM). Avidin binding mode of the ligand molecules has been discussed in the light of docking results. The induced CD profile of the thiazole derivative has been correlated with the stereochemistry of its docked conformation. The important role in the ligand binding of a polar side-chain cluster at the bottom of the biotin-binding cavity as well as the analogous avidin-binding mode of HABA and fenamic acid type NSAIDs have been proposed.
Department of Molecular Pharmacology, Institute of Biomolecular Chemistry, Chemical Research Center, H-1025, Budapest, Pusztaszeri út 59-67, Hungary. zsferi@chemres.hu
Ferenc
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Zsila
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry*; D001360:Avidin / Q000737:chemistry*; D001391:Azo Compounds / Q000737:chemistry; D001665:Binding Sites; D001710:Biotin / Q000737:chemistry*; D002645:Chickens; D002942:Circular Dichroism / Q000379:methods; D008024:Ligands; D015394:Molecular Structure; D011485:Protein Binding; D013844:Thiazoles / Q000737:chemistry; D062367:ortho-Aminobenzoates / Q000737:chemistry
D000894:Anti-Inflammatory Agents, Non-Steroidal; D001391:Azo Compounds; D008024:Ligands; D013844:Thiazoles; D062367:ortho-Aminobenzoates; D001360:Avidin; C001413:HABA; D001710:Biotin; C001209:fenamic acid