Purification of mouse brain ornithine decarboxylase reveals its presence as an inactive complex with antizyme.

Mouse brain ornithine decarboxylase (ODC) was purified to near-homogeneity by using (NH4)2SO4 precipitation and chromatography on heparin-Sepharose, pyridoxamine phosphate-agarose and DEAE-cellulose. On SDS/polyacrylamide-gel electrophoresis, the final preparation gave one protein band similar to that obtained for purified mouse kidney enzyme, corresponding to an Mr of 53.000. The overall yield of the purification exceeded about 50-fold the total activity of the enzyme in the starting material. By affinity chromatography on ODC-bound Sepharose, the extra enzyme activity was shown to originate, at least partly, from the enzyme-antizyme complex. These results demonstrate that ODC in mouse brain occurs mainly in an inactive form and is activated during purification.

Multiple species of ornithine decarboxylase in mouse kidney. Effect of testosterone.

Multiple species of ornithine decarboxylase were separated by chromatography of mouse kidney extract on DEAE-Sepharose CL-6B. The elution patterns of ornithine decarboxylase activity and immunoreactive enzyme protein in the kidneys of untreated and testosterone-treated male mice did not differ otherwise than in order of magnitude. The immunoblots of the chromatography fractions neither revealed any differences in enzyme subunit size between two experimental groups. These findings suggest that the stabilization of ornithine decarboxylase by androgens is not due to the molecular changes of enzyme protein.

Possible involvement of humoral regulation in the effects of elevated cerebral 4-aminobutyric acid levels on the polyamine metabolism in brain.

It has been reported in several recent studies that the manipulation of cerebral 4-aminobutyric acid (GABA) level results in unexpected changes in the cerebral polyamine metabolism in vivo. The mechanisms behind these interactions have remained unknown. The present results show that the changes in polyamine metabolism are not limited to the brain, but are observable also in the liver, which served as a peripheral reference tissue. Different types of responses in the activities of the polyamine-synthesizing enzymes, ornithine decarboxylase and adenosylmethionine decarboxylase, were observed after increasing the cerebral GABA concentration of mice with varying doses of two GABA transaminase inhibitors, gabaculine and ethanolamine-O-sulphate. The time course of the significant changes in the enzyme activities showed significant correlation between the brain and liver. The possibility of direct effects of the drugs on liver was excluded by injecting them intracerebroventricularly, and by performing control experiments with equal doses given peripherally. It is concluded that the observed changes in the polyamine metabolism of liver are produced through centrally mediated humoral regulation, and that the corresponding changes in the brain are obviously due to the same factor or factors, since they are significantly correlated to the changes in liver.

On the metabolism of ornithine in synaptosomal preparations.

The present results show that ornithine is metabolized to glutamate by isolated synaptosomes from mouse cerebral cortex. Under the experimental conditions used the glutamate was channelled further to the tricarboxylic acid cycle, and to a lesser degree to GABA. The possible significance of these metabolic pathways are discussed. Results of an earlier study suggest an excessive metabolism of ornithine via putrescine to GABA in synaptosomes. Those results could not be verified in the present study and a possible reason for the disagreement is demonstrated. However, the present results suggest that putrescine, which is known to be produced from ornithine elsewhere in the nervous tissue, may be metabolized to GABA in synaptosomes.