Abstract
Rabbit muscle phosphorylase phosphatase (E.C. 3.1.3.17) has been purified 6000-fold by isolation of a glycogen-protein complex to which a large part of the enzyme is bound, disruption of the complex with α-amylase then affinity-chromatography of the resulting solution on a Sepharose-polylysine column followed by elution with 6 M urea. The purified enzyme has a specific activity of 2 μmoles of phosphorylase a converted/min/mg. It is free of bound phosphate, carbohydrates, lipids, nucleic acids; it is also free of other enzymatic activities, particularly of unspecific phosphatases. The purified enzyme does not appear to be homogeneous by polyacrylamide gel electrophoresis, probably due to its strong tendency to aggregate. The minimum molecular weight of the active species is 32,000. It has an absolute requirement for SH groups and is resistant to proteolytic enzymes and 8 M urea. The phosphatase activity is strongly dependent upon the state of aggregation of its substrate phosphorylase a and, in most instances, the changes observed are due to a Vm rather than a Km effect. Previous results suggesting a regulation of the phosphatase activity in a glycogen-protein complex have been reinvestigated in the light of the observations described herein.
Supported by grants from the National Institutes of Arthritis and Metabolic Diseases (AM 07902), NIH, USPHS, The National Science Foundation (GB 20482) and the Muscular Dystrophy Association of America.
On leave of absence from the Centre National de la Recherche Scientifique (France).
Supported in part by USPHS grant #HL 10099.
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© 1974 Springer-Verlag Berlin · Heidelberg
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Gratecos, D., Detwiler, T., Fischer, E.H. (1974). Purification and Properties of Rabbit Muscle Phosphorylase Phosphatase. In: Metabolic Interconversion of Enzymes 1973. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-80817-3_5
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DOI: https://doi.org/10.1007/978-3-642-80817-3_5
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