Characterization of highly purified dopamine beta-hydroxylase.
A modified purification procedure has been developed for dopamine beta-hydroxylase isolated from bovine adrenal medulla. Catalase is included in the homogenization step starting with a suspension of either chromaffin granules or adrenal medulla tissue. With this precaution, the enzyme remains stable in the supernatant solution in preparation for the subsequent purification step involving concanavalin A-Sepharose chromatography. The homogeneous enzyme has a specific activity in the range of 60-70 mumol O2 consumed/min/mg. Using radiolabeled metal ion chelators, it was determined that several of the chelators remained tightly bound to the enzyme after removal of the copper leading to difficulties in establishing stoichiometry of enzyme-bound metal ions.
D000313:Adrenal Medulla / Q000201:enzymology*; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002374:Catalase / Q000494:pharmacology; D002417:Cattle; D002837:Chromaffin Granules / Q000201:enzymology*; D002838:Chromaffin System / Q000201:enzymology*; D002845:Chromatography; D003300:Copper / Q000378:metabolism; D004299:Dopamine beta-Hydroxylase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D004492:Edetic Acid / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D007700:Kinetics; D008314:Malonates / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism
D008314:Malonates; D006639:Histidine; D003300:Copper; D004492:Edetic Acid; C030290:malonic acid; D002374:Catalase; D004299:Dopamine beta-Hydroxylase; D001205:Ascorbic Acid; D010100:Oxygen