title: [Electron paramagnetic resonance study of the interactions between steroid hormones and binding proteins].
abstract: Interaction of a spin labeled corticosteroid (desoxycorticosterone nitroxyde: DOC -NO) with three purified proteins (albumin, transcortin, progesterone binding protein: PBG) was studied by electron spin resonance (ESR) spectroscopy. DOC-NO was competitive with natural corticosteroids and therefore bound at the same site to specific binding proteins. ESR spectra in the presence of each of the proteins showed an immobilized (bound) form of the spin labeled steroid and allowed the calculation of the corresponding association constant (Ka) at equilibrium. The three binding proteins could be characterized by the ESR parameters of the DOC-NO bound form. The thermodynamic parameters (deltaH, deltaS) of the steroid-protein interactions were calculated from the ESR data obtained within a wide temperature range (3--40 degrees C). The ESR spectra width (2T) was used to evaluate the polarity of the spin label environment within the steroid binding site: a hydrophobic character was observed for transcortin whereas PBG exhibited a more hydrophilic steroid binding sits. The rotational correlation time of the three protein DOC-NO complexes at equilibrium were calculated from ESR data; the results were correlated with the protein molecular size and suggested a non spherical shape for the binding macromolecule in solution. Spin labelling of biologically active steroids thus provides a novel approach for the study of the interaction of these hormones with their binding protein. Providing a suitable spin label, the ESR parameters may allow the characterization of several types of binding sites of different biological significance for the same hormone, in biological fluids as well as in target tissues.
mesh_terms: D000510:Alpha-Globulins / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism*; D003900:Desoxycorticosterone / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D006728:Hormones / Q000378:metabolism; D006801:Humans; D011487:Protein Conformation; D012399:Rotation; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013696:Temperature; D013816:Thermodynamics; D014156:Transcortin / Q000378:metabolism
chemical_list: D000510:Alpha-Globulins; D002352:Carrier Proteins; D006728:Hormones; D012710:Serum Albumin, Bovine; D003900:Desoxycorticosterone; D014156:Transcortin
vernacular_title: Etude en résonance paramagnétique électronique (RPE) des interactions stéroïdes hormonaux-protéines de liaison.