Steroid-protein interactions. Stopped flow fluorescence studies of the interaction between steroid hormones and progesterone-binding globulin.
Stopped flow fluorometry, measuring changes in the intrinsic fluorescence of progesterone-binding globulin (PBG), was used to determine the association and dissociation rates of the interaction of PBG with seven delta4-3-ketosteroids. The rates of formation and dissociation of the PBG-progesterone complex were measured as a function of concentration and temperature. At 20 degrees, kon = 8.7 X 10(7) M-1 S-1 and koff = 0.060 S-1. The association rate constants for progesterone, deoxycorticosterone, testosterone, testosterone acetate, and medrogestone were found to be the same within experimental error. The different affinities of PBG for these steroids result from the dissociation rate constants of the steroids which ranged from 0.43 S-1 for testosterone to 0.024 S-1 for medrogestone. Two corticosteroids, corticosterone and cortisol, were both bound somewhat more slowly (approximately 5 X 10(7) M-1 S-1). Reflecting their very low affinity for PBG both steroids dissociate very rapidly: corticosterone at 1.4 S-1 and cortisol at 90 S-1. The ratio of association to dissociation rate constants gave affinity constants in agreement with independently determined constants.
D000818:Animals; D001665:Binding Sites; D003345:Corticosterone / Q000097:blood; D003900:Desoxycorticosterone / Q000097:blood; D005260:Female; D006168:Guinea Pigs; D006728:Hormones / Q000097:blood*; D006854:Hydrocortisone / Q000097:blood; D007700:Kinetics; D008524:Medrogestone / Q000097:blood; D011247:Pregnancy; D011374:Progesterone / Q000097:blood*; D011485:Protein Binding; D011956:Receptors, Cell Surface; D012712:Serum Globulins; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013739:Testosterone / Q000097:blood
D006728:Hormones; D011956:Receptors, Cell Surface; D012712:Serum Globulins; D008524:Medrogestone; D013739:Testosterone; D003900:Desoxycorticosterone; D011374:Progesterone; D003345:Corticosterone; D006854:Hydrocortisone