Denaturation of proteins by ascorbic acid: Effects on dopamine-?-hydroxylase

Abstract

Under conditions that are optimum for DbetaH, ascorbic acid denatures serum albumin, gamma-globulin, catalase, and DbetaH. With ascrobate plus Cu2+, the proteins are almost completely destroyed. Pyrazole protects DbetaH and albumin, but not catalase. Superoxide dismutase (SOD) is not denatured by ascorbate, with or without Cu2+, and in combination with catalytic amounts of catalase or Fe2+ it stimulates maximum DbetaH activity. In other words, a combination of catalase and SOD, or Fe2+ and SOD, will protect DbetaH. Excessive amounts of catalase and/or other protein, either native or denatured will prevent the effects of superoxide and/or ascrobate, but cannot replace the requirements for catalytic quantities of catalase or Fe2+. The results suggest that the rate of hydroxylation of tyramine may be limited by superoxide, but that the latter per se does not denature DbetaH as does hydrogen peroxide. The in vitro activation of oxygen by DbetaH is a toxic process, involving the production of both hydrogen peroxide and superoxide and possibly other free radicals. In the absence of precise regulation of the production and concentrations of these compounds, the enzyme is denatured.

Localization of lysophosphatidylcholine in bovine chromaffin granules

“…In this respect it is worth mentioning that the lysophospholipase activity remained constant throughout the incubation period. Thus, enzyme inactivation, due either to deficient trypsin inhibition or to granular ascorbic acid autooxidation [41,42] is not the cause of the incomplete lysophosphatidylcholine hydrolysis in ghosts.…”