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Testosterone Replacement, Low T, HCG, & Beyond
Testosterone and Men's Health Articles
The Dynamics of Allosteric Binding of Estradiol to SHBG
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<blockquote data-quote="madman" data-source="post: 199555" data-attributes="member: 13851"><p><strong>Figure 1. The equilibrium dialysis experiments demonstrate that estradiol binding to the SHBG dimer is a dynamic, nonlinear process. Panel A displays the binding isotherm, generated by the titration of increasing SHBG concentrations in the presence of a fixed estradiol concentration (either 2.6 or 13.7 nM). The data from representative experiments conducted at the respective estradiol concentrations of either 2.6 nM (blue circles) or 13.7 nM (red squares) are shown. The binding isotherm shows nonlinearity of binding as well as asymmetry around the 50% [BE2] point which is inconsistent with the prevalent notion of linear binding with a single dissociation constant, Kd. Panel B depicts the corresponding depletion curve, generated by plotting the concentration of the free estradiol that was dialyzed into the buffer side of the dialysis chamber against the SHBG concentration. Panel C displays the fraction of estradiol that was bound to SHBG ([bound E2] / [total E2]) on the sample side of the dialysis chambers at each SHBG concentration. Each data point represents the average E2 and SHBG concentration from two experiments. Panel D displays the fraction of estradiol that was free ([free E2] / [total E2]) at each SHBG concentration. Each data point represents the average of free E2 and SHBG concentration from two experiments. Panel E is a plot of the apparent Kd vs the SHBG concentration in the equilibrium dialysis experiments performed at five E2 concentrations: 2.6 nM (blue circles), 3.8 nM (green diamonds), 4.1 nM (black triangles), 7.1 nM (purple inverted triangles), and 13.7 nM (red squares). At each SHBG concentration, the apparent Kd was determined from the measured SHBG, and the measured free and bound estradiol concentrations. The apparent Kd varied non-linearly as the SHBG concentrations and SHBG: E2 ratios were varied.</strong></p><p><strong>[ATTACH=full]13799[/ATTACH]</strong></p></blockquote><p></p>
[QUOTE="madman, post: 199555, member: 13851"] [B]Figure 1. The equilibrium dialysis experiments demonstrate that estradiol binding to the SHBG dimer is a dynamic, nonlinear process. Panel A displays the binding isotherm, generated by the titration of increasing SHBG concentrations in the presence of a fixed estradiol concentration (either 2.6 or 13.7 nM). The data from representative experiments conducted at the respective estradiol concentrations of either 2.6 nM (blue circles) or 13.7 nM (red squares) are shown. The binding isotherm shows nonlinearity of binding as well as asymmetry around the 50% [BE2] point which is inconsistent with the prevalent notion of linear binding with a single dissociation constant, Kd. Panel B depicts the corresponding depletion curve, generated by plotting the concentration of the free estradiol that was dialyzed into the buffer side of the dialysis chamber against the SHBG concentration. Panel C displays the fraction of estradiol that was bound to SHBG ([bound E2] / [total E2]) on the sample side of the dialysis chambers at each SHBG concentration. Each data point represents the average E2 and SHBG concentration from two experiments. Panel D displays the fraction of estradiol that was free ([free E2] / [total E2]) at each SHBG concentration. Each data point represents the average of free E2 and SHBG concentration from two experiments. Panel E is a plot of the apparent Kd vs the SHBG concentration in the equilibrium dialysis experiments performed at five E2 concentrations: 2.6 nM (blue circles), 3.8 nM (green diamonds), 4.1 nM (black triangles), 7.1 nM (purple inverted triangles), and 13.7 nM (red squares). At each SHBG concentration, the apparent Kd was determined from the measured SHBG, and the measured free and bound estradiol concentrations. The apparent Kd varied non-linearly as the SHBG concentrations and SHBG: E2 ratios were varied. [ATTACH type="full"]13799[/ATTACH][/B] [/QUOTE]
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Testosterone Replacement, Low T, HCG, & Beyond
Testosterone and Men's Health Articles
The Dynamics of Allosteric Binding of Estradiol to SHBG
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