Protein Phosphatase 2A, Serine, Threonine (PP2A Ser/Thr)

Type 2 protein phosphatases are serine/threonine phosphatases which can preferentially dephosphorylate the a subunit of phosphorylase kinase and are insensitive to inhibitor 2 treatment. Protein phosphatase 2A (PP2A) can be distinguished from PP2B and PP2C which require Ca2+ and Mg2+, respectively, by its partial activity in the absence of divalent cations. PP2A activity is potently inhibited by treatment with okadaic acid and shows much stronger activity toward histone phosphorylated by Protein Kinase C and casein phosphorylated by Protein Kinase A than other protein phosphatases.\n\nEnzyme Dilution Buffer (Included):\n(P9107-33D1): Protein Phosphatase 2A, Serine, Threonine (PP2A Ser/Thr), Supplied as a liquid in 20mM MOPS, pH 7.4, 100mM sodium chloride, 60mM 2-mercaptoethanol, 1mM MgCl2, 1mM EGTA, 0.1mM MnCl2, 1mM DTT, 0.1mg/ml serum albumin, 10% glycerol.\n\nApplications:\nSuitable for use in selective in vitro dephosphorylation of proteins. Other applications not tested.\n\nRecommended Dilutions:\nSelective in vitro dephosphorylation: Use 0.05-0.5u total of PP2A per reaction. Note: Phosphatase activity is inhibited by 10nM okadaic acid. Some preparations show extremely low levels of serine kinase activity (1-3fmole/min/mg), which can be abolished if needed by the addition of EDTA to the assay buffer.\n\nUnit Definition: \nOne Unit releases 1nmole of phosphate per min from 15uM [32P] labeled Phosphorylase A at 30 degrees C.\n\nStorage and Stability:\nAliquot to avoid repeated freezing and thawing.. 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. \n\nDO NOT STORE AT -70C

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SPECIFICATIONS

Catalog Number

P9107-33D

Size

10U

Form

Supplied as a liquid in, 20mM MOPS, pH 7.5, 500mM sodium chloride, 60mM 2-mercaptoethanol, 1mM MgCl2, 1mM EGTA, 0.1mM MnCl2, 1mM DTT, 0.1mg/ml serum albumin, 10% glycerol.

Purity

Purified by a combination of DEAE, aminohexyl-agarose and gel filtration chromatography. Isolated as the heterodimer of 60kD (A) and 36kD (C) subunits.

References

1. Ito, et al., EMBO J. 19: 562-571 (2000). 2. Sathyanarayanan, S., et al., Cell 116: 603-615 (2004). 3. Usui, et al., J. Biol. Chem. 258: 10,455 (1983). 4. Shenolikar and Nairn, Adv. Sec. Messenger Phosphoprotein Res. 23: 1 (1990). 5. Anderson, et al., Nature 1343: 651 (1991). 6. DeFranco, et al., Mole. Endocrinol. 5: 1215 (1991). 7. Reinhart, et al., J. Neurosci. 11: 1627 (1991). 8. Scheidtmann, Mole. Cell Biol. 11: 1996 (1991).

Additional Info

Specific for p-Ser and p-Thr proteins as compared to p-Tyr. Reported preference for sites in signal transduction kinases such as MAP kinases, steroid receptor, ion channels and p53 protein. This PP2A is highly active and displays characteristic properties of Type-2A phosphatases. Reconstitutes to ABC trimer with added B subunits. Extremely sensitive to okadaic acid (inhibition at < 10nM) and resistant to inhibitor-2. Phosphate, phosphoesters and fluoride are inhibitory. Sulfhydryl compounds are required to maintain activity.