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Affinity Chromatography of Regulatory Subunits of Protein Phosphatase-1
- Source :
- Archives of Biochemistry and Biophysics. 325:82-90
- Publication Year :
- 1996
- Publisher :
- Elsevier BV, 1996.
-
Abstract
- In this study we demonstrate that recombinant rabbit muscle protein phosphatase-1 immobilized on CH-Sepharose is an efficient affinity chromatography support for the isolation of subunits of phosphatase-1. The support was used to isolate the glycogen binding subunit of phosphatase-1 from muscle and nonmuscle rat tissues. Examination of the affinity-purified material from rat heart and liver showed that the major component was a 160-kDa polypeptide on SDS-PAGE. The identity of the purified liver 160-kDa polypeptide as the glycogen binding subunit was confirmed by the demonstration that it is capable of binding to glycogen, and is phosphorylated by the catalytic subunit of PKA. The novel observation was made that the phosphorylation was dependent on the presence of glycogen. Examination of the material from heart, lung, liver, kidney, and brain showed a similar phenomenon. Our studies show that this subunit is widely distributed in tissues. The affinity support was also efficient in the isolation of the NIPP-1 (nuclear inhibitor of protein phosphatase-1) proteins from calf thymus. Examination of heat-treated extracts of rat liver led to the isolation of a novel 19-kDa inhibitory protein which could also be phosphorylated by PKA and may represent the rat liver homolog of calf thymus NIPP-1.
- Subjects :
- Protein subunit
Immunoblotting
Biophysics
Thymus Gland
Biology
Biochemistry
Chromatography, Affinity
law.invention
chemistry.chemical_compound
Affinity chromatography
law
Protein Phosphatase 1
Phosphoprotein Phosphatases
Animals
Enzyme Inhibitors
Phosphorylation
Muscle, Skeletal
Molecular Biology
chemistry.chemical_classification
Glycogen binding
Glycogen
Myocardium
Intracellular Signaling Peptides and Proteins
Proteins
Protein phosphatase 1
Enzymes, Immobilized
Cyclic AMP-Dependent Protein Kinases
Molecular biology
Rats
Enzyme
Liver
chemistry
Organ Specificity
Recombinant DNA
Cattle
Electrophoresis, Polyacrylamide Gel
Rabbits
Carrier Proteins
Subjects
Details
- ISSN :
- 00039861
- Volume :
- 325
- Database :
- OpenAIRE
- Journal :
- Archives of Biochemistry and Biophysics
- Accession number :
- edsair.doi.dedup.....5b5e28be4bb7984f94c7cfbef1a2e0c1