J. Biochem, 1981, Vol. 89, No. 2 609-614
© 1981 Japanese Biochemical Society
research-article |
Succinyl Trialanine p-Nitroanilide-Hydrolytic Enzymes in Human Serum. Partial Purification and Characterization
*Department of Biochemistry, Nagoya City University Medical School
**Department of Internal Medicine, Nagoya City University Medical School, Mizuho-ku, Nagoya, Aichi 467
The levels of two kinds of elastase-like enzymes, which are able to hydrolyze an artificial elastase substrate, suc-(Ala)3-pNA, but unable to hydrolyze a naturally occurring substrate, elastin, were found to be elevated in the sera of patients suffering from hepatobiliary disorders and other diseases accompanied by tissue damage. One of the enzymes was characterized as being sensitive to a chelating reagent, EDTA, and partially inactivated enzyme activity was recovered by the addition of calcium ion. The apparent molecular weight estimated by Sepharose 4B column chromatography showed a wide distribution from 200,000 to approximately 10,000,000, but all components were converted to a molecular weight of about 200,000 by treatment with 2% Triton X-100. The activity of this enzyme was partially reduced by the addition of anti-ß-lipoprotein antibody, showing that a part of the enzyme was affiliated with low and very low density lipoproteins in the serum. The level of the other enzyme was rarely increased in the sera of patients suffering from severe hepatic disorders. This enzyme was resistant to EDTA, and the apparent molecular weight was 150, 000–200, 000. It appeared not to be associated with lipid component. Both enzymes were assumed to be tissue-derived enzymes, because their activities were very low in the sera of healthy persons.