| Abstract: | 1) A lysosomal protease, a new cathepsin that inactivates glucose-6-phosphate dehydrogenase EC 1.1.1.49] and some other enzymes and differs from cathepsin B EC 3.4.22.1] was purified about 2,200-fold from crude extracts of rat liver by cell-fractionation, freezing and thawing, acetone treatment, gel filtration, and DEAE Sephadex and CM-Sephadex column chromatographies. 2) The new cathepsin was markedly activated by the thiol-reagent, 2-mercaptoethanol and inhibited by monoiodoacetate. 3) The molecular weight of the new cathepsin was found by Sephadex G-75 column chromatography to be 22,000, which is smaller than that of cathepsin B. 4) The optimum pH of the enzyme for inactivation of glucose-6-phosphate dehydrogenase was pH 5.0--5.5. The enzyme was unstable in alkali and on heat treatment. 5) The rates of inactivation of glucose-6-phosphate dehydrogenase, apo-ornithine aminotransferase EC 2.6.1.13], apo-tyrosine aminotransferase EC 2.6.1.5], apo-cystathionase EC 4.4.1.1], glucokinase EC 2.7.1.2], glyceraldehyde-3-phosphate dehydrogenase EC 1.2.1.12], and malate dehydrogenase EC 1.1.1.37] by the new cathepsin were higher than those by cathepsin B. However aldolase EC 4.1.2.13] was inactivated more rapidly by cathepsin B than by the new cathepsin. Lactate dehydrogenase EC 1.1.1.27], glutamate dehydrogenase EC 1.4.1.2] and alcohol dehydrogenase EC 1.1.1.1] were not inactivated by either cathepsin. Unlike cathepsin B, the new cathepsin scarcely hydrolyzes N-substituted derivatives of arginine. |
