Portal de Periódicos da CAPES

Application of general ligand affinity chromatography for the mutual separation of deoxyribonuclease and ribonuclease free of protease contamination

1976; Elsevier BV; Volume: 74; Issue: 1 Linguagem: Inglês

10.1016/0003-2697(76)90317-1

1096-0309

L.H. Lazarus, C.-Y. Lee, Bendicht Wermuth,

Enzyme function and inhibition

Abstract α-Chymotrypsin and trypsin could be removed from mixtures of pancreatic RNase and DNase I by chromatography on the general ligand affinity column of 8-(6-aminohexyl)-amino-5′-AMP-Sepharose. Both RNase and DNase are quantitatively recovered by elution with 5′-AMP. The separation of RNase and DNase from each other was carried out by the specific retention of RNase on 8-(6-aminohexyl)-amino-2′-AMP-Sepharose at high ionic strength (0.1 m ); DNase failed to adsorb under these conditions. RNase was then desorbed in the presence of a mononucleotide. This is a facile method for obtaining pure enzymes from commercial preparations and gives rise to the potential application of these columns for the analysis of enzyme-nucleotide interactions.