Purification of 2',5'-oligoadenylate synthetase from rabbit reticulocytes.
The 2',5'-oligoadenylate synthetase (2-5A synthetase) from rabbit reticulocytes has been purified to apparent homogeneity. The purification procedure consists of (NH4)2SO4 fractionation (30-50% cut), specific binding of the 2-5A synthetase to and elution from the affinity matrix of polyinosinic-polycytidylic-cellulose, another (NH4)2SO4 precipitation step, and finally chromatography on DEAE-cellulose. Upon electrophoresis in sodium dodecyl sulfate polyacrylamide gel (10%), the purified enzyme migrates as a single polypeptide with an apparent molecular weight of 110,000 daltons. A sedimentation coefficient of 5.8S is obtained by glycerol density gradient centrifugation. The synthesis of 2',5'-oligoadenylate by the purified enzyme is dependent on the presence of double-stranded (ds) RNA, in the absence of which the enzyme is highly unstable. Biochemical characteristics of the purified enzyme have been defined.
D015088:2',5'-Oligoadenylate Synthetase / Q000097:blood* / Q000302:isolation & purification; D000818:Animals; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D007700:Kinetics; D008970:Molecular Weight; D011070:Poly I-C; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*
D015088:2',5'-Oligoadenylate Synthetase; D011070:Poly I-C