Purification and characterization of cathepsin D-like proteinase from the tadpole tail of bullfrog, Rana catesbeiana.
1. An acid aspartic proteinase in the regressing tadpole tail was purified about 800-fold with a 36% recovery. 2. The mol. wt of the enzyme was found to be 42,000 on gel filtration and 38,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively. 3. The purified enzyme had a maximum activity at pH 3.5 and an apparent Km of 0.084% with acid-denatured hemoglobin as substrate. 4. The enzyme activity was strongly inhibited by pepstatin. In addition, diazoacetylnorleucine methyl ester inactivated the enzyme in the presence of cupric ions. 5. The enzyme was identified as a cathepsin D (EC. 3.4.23.5)-like proteinase.
Nanbu
M
M
Life Science Institute, Sophia University, Tokyo, Japan.
Kobayashi
K
K
Horiuchi
S
S
D000818:Animals; D002402:Cathepsin D / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007814:Larva / Q000201:enzymology; D008970:Molecular Weight; D010436:Pepstatins / Q000494:pharmacology; D011892:Rana catesbeiana / Q000378:metabolism*; D013379:Substrate Specificity; D013623:Tail
D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D002402:Cathepsin D; C031375:pepstatin