Epidermal nucleases: purification and characterization of ribonuclease from mammalian epidermis.
The major ribonuclease of adult guinea pig epidermis has been isolated and purfied over 1000-fold by a combination of ammonium sulfate fractionation, affinity and ion-exchange chromatography, and electrophoresis. The purified enzyme is free from phosphodiesterase and phosphatase activities. The ribonuclease is optimally active near neutrality in phosphate buffer, with a Km of 3mu g/ml toward [14-C]RNA from Erhlich ascites tumor cells. (here are no metal requirements for activity. The enzyme catalyzes the endonucleolytic hydrolysis of high molecular weight yeast RNA and it also hydrolyzes polycytidylic and polyuridylic acids, but not polyadenylic, polyguanylic, and polyinosinic acids. The apparent molecular weight of the active enzyme is 28 500.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002286:Carcinoma, Ehrlich Tumor; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D004589:Electrophoresis, Disc; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011119:Polynucleotides; D011188:Potassium / Q000494:pharmacology; D012334:RNA, Neoplasm; D012260:Ribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D012867:Skin / Q000201:enzymology*; D012964:Sodium / Q000494:pharmacology; D013696:Temperature
D011119:Polynucleotides; D012334:RNA, Neoplasm; D012964:Sodium; D012260:Ribonucleases; D010744:Phosphoric Monoester Hydrolases; D010727:Phosphoric Diester Hydrolases; D008274:Magnesium; D011188:Potassium; D002118:Calcium