Protein Phosphatase 1, Ser/Thr (PP1)

This catalytic fragment of PP1 is highly active and displays properties characteristic of type-1 phosphatases:3 high specific activity with phosphorylase as substrate, relative resistance to okadaic acid (up to 50nM), and sensitivity to inhibitor-2. Sulfhydryl compounds are required to maintain activity. Phosphate, phosphoesters, and concentrated salts are inhibitory. \n\nApplications:\nSuitable for use In vitro dephosphorylation of proteins. Other applications not tested.\n\nRecommended Dilution:\nIn vitro dephosphorylation:\n0.1-0.5 units total PP1 per reaction. A unit releases 1nM of phosphate/min from 15uM phosphorylase at 30 degrees C. Inhibition by 50nM microcystin-LR or 500nM okadaic acid has been demonstrated previously. The dilution buffer should be made 50% with glycerol if diluted solutions are to be stored at -20 degrees C. The activity was determined by a malachite green based phosphatase assay.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nFor long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P9107-05

Size

10U

Form

Supplied as a liquid in 100ul of 50% glycerol, 20mM MOPS, pH 7.5, 60mM 2-mercaptoethanol, 1mM EGTA, 0.1mM MnCl2, 1mM MgCl2 and 0.1mg/ml serum albumin.

Purity

Purified by a combination of DEAE chromatography, divalent cation activation plus trypsin digestion, and gel filtration chromatography.1 A mixture of PP1 isoforms are present, based on peptide sequencing.

References

1. Brautigan, et al., J. Biol. Chem. 260: 4295, 1985. 2. Johnson, et al., Mol. Endocrinol. 1: 745, 1987. 3. Shenolikar and Nairn, Adv. Sec. Messenger Phosphoprotein Res. 23: 1, 1990. 4. Hagiwara, Cell 70: 105, 1992. 5. DeFranco, et al., Molec. Endocrinol. 5: 1215, 1991. 6. Chung, et al., Science 253: 560, 1991. 7. Ingebritsen and Cohen, Eur. J. Biochem. 132: 255, 1983.

Additional Info

Absolute specificity for protein P-Ser and P-Thr compared to P-Tyr. Reported preference for sites in transcription factor,4 steroid receptor,5 ion channels, 6 and enzymes of glycogen metabolism.