Analysis of the kinetics of ovine follitropin agonist-antagonist interactions with pig ovarian membranes.
The binding of 125I-labeled ovine follitropin (oFSH) and 125I-labeled deglycosylated ovine follitropin (DG-oFSH) to porcine granulosa cell membranes was studied at equilibrium and nonequilibrium binding conditions and statistically analyzed. Saturation and competition binding experiments revealed homogeneity in the population of binding sites labeled with 125I-oFSH, having a pK estimation of approximately equal to 10. 125I-DG-oFSH similarly interacts with a single uniform class of receptors of equal affinity (pK approximately equal to 10) and binding capacity as oFSH. In contrast, displacement experiments using 125I-DG-oFSH as tracer and unlabeled oFSH as competing ligand demonstrate slope factors less than unity, suggesting apparent heterogeneity of sites not observed with 125I-DG-oFSH vs. DG-oFSH competition experiments. Under these conditions, it appears that FSH binds to two sites in near equal proportion but of unequal affinities. The total specific binding capacities of these sites equal those observed in 125I-DG-oFSH/unlabeled DG-oFSH competition experiments. Analysis of oFSH association kinetics at 37 degrees C by curve-fitting methods is best explained by a biexponential rate equation describing a fast and a slow association component that are equally distributed. DG-oFSH demonstrates a disproportionately greater amount of fast vs. slow binding component. The binding half-times for each component of oFSH and DG-oFSH are similar, i.e., minutes for the fast and hours for the slow t 1/2 times. At 37, 25, and 4 degrees C, DG-oFSH exhibits greater velocity of binding to the receptor than oFSH.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000031:analogs & derivatives / Q000378:metabolism*; D006107:Granulosa Cells / Q000378:metabolism*; D007700:Kinetics; D011962:Receptors, FSH / Q000378:metabolism*; D012756:Sheep; D013552:Swine
D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone