Occurrence and Some Properties of Membrane-Bound Neutral Proteinase in the Microsomal Fraction of Rat Skeletal Muscle1
| Autor: | Yoshikazu Ichihara, Kazuhiro Sogawa, Kenji Takahashi |
|---|---|
| Rok vydání: | 1982 |
| Předmět: | |
| Zdroj: | The Journal of Biochemistry. 91:87-95 |
| ISSN: | 1756-2651 0021-924X |
| Popis: | Membrane-bound neutral proteinase was found in the microsomal fraction of rat skeletal muscle as assayed with heat-denatured casein as a substrate. The enzyme was solubilized from 1 M KCl-washed microsomal fraction by 1 % sodium cholate containing 0.1 M NaCl, and partially purified by chromatograph y on a column of Sepharose CL-6B in the presence of 0.5% sodium cholate and 0.1 M NaCl. The enzyme was eluted from the Sepharose column as a single but rather broad peak at a position corresponding to a molecular weight of about 190,000. The pH optimum for hydrolysis of heat-denature d casein was about 8.0. It was inhibited to significant extents by various reagents including diisopropyl phosphorofluoridate, phenylmethanesulfonyl fluoride, iVMosyl-L-phenylalanine chloromethyl ketone, iVMosylL-lysine chloromethyl ketone, p-chloromercuriphenyl sulfonate, chymostatin, EDTA, EGTA, and o-phenanthroline. This inhibition profile suggests that the present muscle proteinase is a mixture of proteinases, such as a serine proteinase and a metallo-proteinase similar to those occurring in the microsomal membranes of liver and kidney (or small intestine), respectively. Among urea-denatured proteins tested as substrates, calf thymus histone was hydrolyzed most rapidly, followed by protamine, hemoglobin, and casein. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|