Conformational Changes of Blood ACE in Chronic Uremia
Figure 3
Blood ACE phenotyping in uremia.
A. ACE activity in the citrated plasma (diluted 5-fold) of patients with uremia (versus that of healthy volunteers) was determined spectrofluorometrically using 20 µl of diluted plasma with 100 µl of ACE substrate ZPHL at 1 hour of incubation with substrate. B. The ratio of the rates of the hydrolysis of two substrates (ZPHL/HHL ratio) for ACE from the blood of patients with uremia and from the blood of healthy persons. C-E. ACE activity precipitated by different mAbs from citrated plasma of uremic patients (versus that from plasma of healthy donors: E - by mAb 9B9; D - by mAb 1G12 and C – their ratio for each plasma sample. Results are shown as mean ± SD from 3 to 10 independent experiments, each in duplicates or triplicates. The red color of the columns shows plasma of those patients whose blood parameter (ACE activity, Z/H ratio, 1G12/9B9 ratio/mAb 9B9 or 1G12 binding) was significantly higher (and yellow bar-significantly lower) than mean value for healthy donors. The green bars shows mean values for healthy controls. Boxes show those patients which have increased 1G12/9B9 ratio while normal ZPHL/HHL ratio. All other terms and conditions are as in Fig. 1. * - p<0.05 in comparison with mean value for healthy controls.