Synthesis of chromogranins and dopamine beta-hydroxylase by perfused bovine adrenal glands.

The incorporation of [3H]leucine into chromogranins and into soluble and membrane-bound dopamine beta-hydroxylase (DBH) was studied in isolated perfused adrenal glands. [3H]chromogranins and [3H]DBH were quantitatively determined by immunoprecipitation. The amounts of soluble [3H]DBH formed were about equal to the amounts of membrane-bound [3H]DBH whereas the amounts of [3H]chromogranin were 5- to 20-fold greater than that of soluble [3H]DBH. On continuous sucrose density gradients, [3H]chromogranin was unimodally distributed after 2- to 20-h chase periods and accumulated in a vesicle having a lower buoyant density than mature chromaffin vesicles. At 2 h both membrane and soluble [3H]DBH were both bimodally distributed whereas after a 20-h chase period the distribution of soluble and membrane [3H]DBH was essentially unimodal and paralleled the distribution of [3H]chromogranin. These studies indicate that [3H]chromogranin, soluble [3H]DBH, and membrane [3H]DBH are synthesized concomitantly, but that each is transported into chromaffin vesicles at different rates.