Abstract
A simple, large scale purification procedure for the violet “phosphoprotein phosphatase” from beef spleen [Glomset, J. and Porath, J., Biochim. Biophys. Acta, 39, 1 (1960)] has been devised. The procedure involves a 24-hr extraction of a tissue homogenate at pH 3, ammonium sulfate fractionation, CM-cellulose chromatography, and Sephadex G-75 chromatography. The highly purified enzyme shows a constant ratio of both enzymatic activity and absorbance at 550 nm to iron concentration. The data strongly indicate that the enzyme contains iron, and moreover, support a 1:1 molar ratio of iron to enzyme.
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