Odorant binding and conformational changes of a rat odorant-binding protein.
Odorant-binding proteins (OBPs) are lipocalins secreted in the nasal mucus of vertebrates, which convey odorants to their neuronal receptors. We compared the binding properties of a recombinant rat OBP (OBP-1F) using a set of six odorants of various chemical structures. We examined the binding properties by both fluorescent probe competition and isothermal titration calorimetry. OBP-1F affinity constants, in the micromolar range, varied by more than one order of magnitude and were roughly correlated to the odorant size. The observed binding stoichiometry was found to be around one odorant per dimer. Using tyrosine differential spectroscopy, the binding of ligand was shown to induce local conformational changes. A three-dimensional structure of OBP-1F, modelled using the known structure of aphrodisin as template, allowed us to suggest the location of the observed structural changes outside of the binding pocket. These results are consistent with one binding site located in one of the two beta-barrels of the OBP-1F dimer and a subtle conformational change correlated with binding of an odorant molecule, which hampers uptake of a second odorant by the other hydrophobic pocket.
Nespoulous
C
C
INRA, Biochimie et Structure des Protéines, 78352 Jouy-en-Josas, France. nespoulo@jouy.inra.fr
Briand
L
L
Delage
M-M
MM
Tran
V
V
Pernollet
J-C
JC
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D019281:Dimerization; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D051381:Rats; D018035:Receptors, Odorant / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013816:Thermodynamics
D008024:Ligands; D018035:Receptors, Odorant; D011994:Recombinant Proteins; C045648:odorant-binding protein