Purification and characterization of adenylate kinase isozymes from rat muscle and liver
| Autor: | Hachiro Nakagawa, Takiko Tamura, Hiroshi Shiraki |
|---|---|
| Rok vydání: | 1980 |
| Předmět: |
Male
Adenylate kinase Biology Isozyme chemistry.chemical_compound medicine Animals Tissue Distribution Isoelectric Point Sodium dodecyl sulfate Polyacrylamide gel electrophoresis Gel electrophoresis Muscles Adenylate Kinase Phosphotransferases Skeletal muscle General Medicine Molecular biology Rats Isoenzymes Molecular Weight Kinetics Isoelectric point medicine.anatomical_structure Liver chemistry Biochemistry Sephadex |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Enzymology. 612:56-66 |
| ISSN: | 0005-2744 |
| DOI: | 10.1016/0005-2744(80)90278-8 |
| Popis: | Isozymes of adenylate kinase (ATP:AMP phosphotransferase, EC 2.7.4.3) were purified from skeletal muscle and liver of rats to essentially homogeneous states by acrylamide gel electrophoresis and sodium dodecyl sulfate gel electrophoresis. The isozyme from muscle was purified by acidification to pH 5.0, and column chromatography on phosphocellulose, Sephadex G-75 and Blue Sepharose CL-6B, while that from liver was purified by column chromatography on Blue Sepharose CL-6B, Sephadex G-75 and carboxymethyl cellulose. By these procedures the muscle isozyme was purified about 530-fold in 29% yield, and the liver isozyme about 3600-fold in 27% yield from the respective tissue extracts. The molecular weights of the muscle and liver isozymes were estimated as about 23 500 and 30 500, respectively, by both sodium dodecyl sulfate gel electrophoresis and molecular sieve chromatography, and no subunit of either isozyme was detected. The isoelectric points of the muscle and liver isozymes were 7.0 and 8.1, respectively. The K m values of the respective enzymes for ATP and ADP were similar, but the K m (AMP) of the liver isozyme was about one-fifth of that of the muscle isozyme. Immunological studies with rabbit antiserum against the rat muscle isozyme showed that the muscle isozyme was abundant in muscle, heart and brain, while the liver isozyme was abundant in liver and kidney. |
| Databáze: | OpenAIRE |
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