Epidermal nucleases. II. The multiplicity of ribonucleases in guinea-pig epidermis.
Ribonuclease activity has been extracted from adult guinea-pig epidermis by sequential homogenization in dilute sodium acetate and sulfuric acid. The extracts were subjected to ammonium sulfate fractionation and to affinity and ion exchange chromatography. Three ribonucleases (I, II, III) were separated from the sodium acetate extract and 6(A, B1, B2, B3, C, D) were isolated from the sulfuric acid extract. The degree of purification varies from 65-fold to 8,700-fold and the apparent molecular weights of the active forms of 8 of the 9 ribonucleases range from 10,000 to 36,500. No phosphodiesterase activity is present in any of the 9 fractions, but there is alkaline phosphatase activity in one (I) and deoxyribonuclease activity in a second (B3). Two of the ribonucleases have acid pH optima (a1, B3), while the others are most active between PHs 6.8 and 7.8. The activity of 4 of the fractions is sensitive to added EDTA (III, A, B2, B3,), but no stimulatory metal ions were found. Low concentrations of the polyamine spermidine enhanced the activity of 3-fractions (III, C, D). Yeast ribonucleic acid is degraded exonucleolytically by 2 fractions (I, A) and endonucleolytically by the remaining 7. In experiments with homopolyribonucleotide substrates, poly U was generally the preferred substrate. Substantial hydrolysis of poly A occurred with 2 fractions (A, B3) and slight hydrolysis of poly G with 2 other fractions (B2, C).
D000818:Animals; D003851:Deoxyribonucleases / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D012260:Ribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D012867:Skin / Q000201:enzymology*; D013095:Spermidine / Q000494:pharmacology
D004492:Edetic Acid; D003851:Deoxyribonucleases; D012260:Ribonucleases; D010744:Phosphoric Monoester Hydrolases; D010727:Phosphoric Diester Hydrolases; D013095:Spermidine