Characterization of highly purified ornithine decarboxylase from rat heart.

A highly purified preparation of heart ornithine decarboxylase was obtained from isoproterenol-treated rats. The molecular and catalytic properties of the cardiac enzyme were investigated. The isoelectric point of the enzyme appeared to be 4.9, and the molecular weight was estimated to be 54000 by SDS-polyacrylamide gel electrophoresis. Under nondenaturing conditions, the molecular weight of the partially purified enzyme was 10000-110000 as determined by gel filtration, whereas a significantly lower (Mr approx. 70000) value was obtained for purified ornithine decarboxylase. Both Km for the substrate and Vmax were affected by the dithiothreitol concentration in the assay mixture. In particular, the Km for ornithine was found to be about 0.09 mM in the presence of 2.9 mM dithiothreitol and appeared to decrease at lower dithiothreitol concentrations. The Km for pyridoxal phosphate was about 0.09 microM; putrescine and lysine inhibited the enzyme competitively, with Ki values of 1.3 and 11.7 mM, respectively. The existence of two different forms of ornithine decarboxylase in cardiac tissue was indicated by DEAE-cellulose chromatography.