Purification and properties of a nuclear DNA endonuclease from HeLa S3 cells
| Autor: | George P. Studzinski, Gary J. Fischman, Muriel W. Lambert |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Cations
Divalent Size-exclusion chromatography Biology Coliphages chemistry.chemical_compound Endonuclease Caffeine Polyacrylamide gel electrophoresis Cell Nucleus Deoxyribonucleases Isoelectric focusing Sodium Sulfhydryl Reagents DNA General Medicine Endonucleases Enzyme assay Molecular Weight Isoelectric point Biochemistry chemistry DNA Viral Potassium biology.protein Cell fractionation HeLa Cells |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Enzymology. 567:464-471 |
| ISSN: | 0005-2744 |
| DOI: | 10.1016/0005-2744(79)90132-3 |
| Popis: | An endonuclease that can act on calf thymus DNA and circular double-stranded phage PM2 DNA has been isolated from HeLa S3 cell chromatin. Approximately 200-fold purification was achieved by a sequence of subcellular fractionation, differential NaCl solubility and chromatography on CM-Sephadex, DEAE-cellulose and hydroxyapatite, and isoelectric focusing. The pH optimum of the enzyme is 7.0 ± 0.5 and the isoelectric point is pH 5.1 ± 0.2 . Divalent cations are necessary for its activity and the enzyme is heat inactivated at 60°C. The enzyme activity is sensitive to caffene and sulfhydryl reacting compounds. The molecular weight, determined by gel filtration and SDS gel electrophoresis, is approx. 22 000. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|