Characterization of norepinephrine binding to plasma proteins of the dog and the rabbit

The binding of 3H-norepinephrine (L-3H-NE, 1.0 X 10(-9) M) to plasma proteins of the dog and the rabbit was studied under controlled conditions. Destruction of NE occurred less rapidly at 22 degrees than at 37 degrees. Binding measured at 22 degrees was equivalent to that at 37 degrees, while binding measured at 0 degree was greater than that at 37 degrees. Therefore, losses of plasma NE were minimized by incubation of samples at 22 degrees for no longer than 30 minutes. L-3H-NE binding was examined in the absence and presence of 10(-9) to 10(-2) M non-labeled L-NE, DL-NE, DL-normetanephrine (NM), DL-epinephrine (E), dopamine (DA), and catechol (C). Specific binding of L-3H-NE varied in the range of NE concentrations (L-3H-NE + non-labeled NE) from 10(-9) M (18.7 +/- 3.1%, rabbit; 25.6 +/- 4.8%, dog) to 10(-6) M (10.8 +/- 3.1%, rabbit; 15.2 +/- 3.6%, dog). Calculated binding constants (KD) were consistent with binding to circulating proteins such as globulins or albumin (4.2 +/- 1.2 X 10(-5) M, rabbit; 5.4 +/- 1.7 X 10(-5) M, dog). In plasma from both species, non-labeled DL-NE, L-NE, E, DA, and C, but not NM (from 10(-9) to 10(-2) M) each significantly displaced L-3H-NE from its binding site in a manner similar to displacement produced by non-labeled NE. The results demonstrate that 1) NE is bound to plasma proteins, although to a lesser extent than had been reported by other investigators; and 2) the binding of catecholamines to plasma proteins may be mediated by the catechol ring.