Browse > Article
http://dx.doi.org/10.5483/BMBRep.2010.43.4.263

Phosphorylation on the PPP2R5D B regulatory subunit modulates the biochemical properties of protein phosphatase 2A  

Yu, Un-Young (Department of Biochemistry, Ewha Womans University School of Medicine)
Ahn, Jung-Hyuck (Department of Biochemistry, Ewha Womans University School of Medicine)
Publication Information
BMB Reports / v.43, no.4, 2010 , pp. 263-267 More about this Journal
Abstract
To characterize the biochemical properties of the PP2A regulatory B subunit, PPP2R5D, we analyzed its phosphorylation sites, stoichiometry and effect on holoenzyme activity. PPP2R5D was phosphorylated on Ser-53, Ser-68, Ser-81, and Ser-566 by protein kinase A, and mutations at all four of these sites abolished any significant phosphorylation in vitro. In HEK293 cells, however, the Ser-566 was the major phosphorylation site after PKA activation by forskolin, with marginal phosphorylation on Ser-81. Inhibitory tyrosine phosphorylation on Tyr-307 of the PP2A catalytic C subunit was decreased after forskolin treatment. Kinetic analysis showed that overall PP2A activity was increased with phosphorylation by PPP2R5D phosphorylation. The apparent Km was reduced from $11.25\;{\mu}M$ to $1.175\;{\mu}M$ with PPP2R5D phosphorylation, resulting in an increase in catalytic activity. These data suggest that PKA-mediated activation of PP2A is enabled by PPP2R5D phosphorylation, which modulates the affinity of the PP2A holoenzyme to its physiological substrates.
Keywords
B subunit; Phosphorylation; PPP2R5D; PP2A; Protein kinase A;
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
Times Cited By SCOPUS : 3
연도 인용수 순위
  • Reference
1 Usui, H., Inoue, R., Tanabe, O., Nishito, Y., Shimizu, M., Hayashi, H., Kagamiyama, H. and Takeda, M. (1998) Activation of protein phosphatase 2A by cAMP-dependent protein kinase-catalyzed phosphorylation of the 74-kDa B'' (delta) regulatory subunit in vitro and identification of the phosphorylation sites. FEBS Lett. 430, 312-316   DOI   ScienceOn
2 Vafai, S. B. and Stock, J. B. (2002) Protein phosphatase 2A methylation: a link between elevated plasma homocysteine and Alzheimer's disease. FEBS Lett. 518, 1-4   DOI   PUBMED   ScienceOn
3 Fukui, R., Svenningsson, P., Matsuishi, T., Higashi, H., Nairn, A. C., Greengard, P. and Nishi, A. (2003) Effect of methylphenidate on dopamine/DARPP signalling in adult, but not young, mice. J. Neurochem. 87, 1391-1401   DOI   ScienceOn
4 Bibb, J. A., Chen, J., Taylor, J. R., Svenningsson, P., Nishi, A., Snyder, G. L., Yan, Z., Sagawa, Z. K., Ouimet, C. C., Nairn, A. C., Nestler, E. J. and Greengard, P. (2001) Effects of chronic exposure to cocaine are regulated by the neuronal protein Cdk5. Nature 410, 376-380   DOI   ScienceOn
5 Virshup, D. M. (2000) Protein phosphatase 2A: a panoply of enzymes. Curr. Opin. Cell. Biol. 12, 180-185   DOI   PUBMED   ScienceOn
6 Janssens, V., Longin, S. and Goris, J. (2008) PP2A holoenzyme assembly: in cauda venenum (the sting is in the tail). Trends. Biochem. Sci. 33, 113-121   DOI   PUBMED   ScienceOn
7 Margolis, S. S., Perry, J. A., Forester, C. M., Nutt, L. K., Guo, Y., Jardim, M. J., Thomenius, M. J., Freel, C. D., Darbandi, R., Ahn, J. H., Arroyo J. D., Wang X. F., Shenolikar, S., Nairn, A. C., Dunphy, W. G., Hahn, W. C., Virshup, D. M. and Kornbluth, S. (2006) Role for the PP2A/B56delta phosphatase in regulating 14-3-3 release from Cdc25 to control mitosis. Cell 127, 759-773   DOI   ScienceOn
8 Stipanovich, A., Valjent, E., Matamales, M., Nishi, A., Ahn, J. H., Maroteaux, M., Bertran-Gonzalez, J., Brami-Cherrier, K., Enslen, H., Corbille, A. G., Filhol O., Nairn A. C., Greengard, P., Heve, D. and Girault, J. A. (2008) A phosphatase cascade by which rewarding stimuli control nucleosomal response. Nature 453, 879-884   DOI   ScienceOn
9 Ahn, J. H., Sung, J. Y., McAvoy, T., Nishi, A., Janssens, V., Goris, J., Greengard, P. and Nairn, A. C. (2007) The B''/PR72 subunit mediates $Ca^{2+}$-dependent dephosphorylation of DARPP-32 by protein phosphatase 2A. Proc. Natl. Acad. Sci. U.S.A. 104, 9876-9881   DOI   ScienceOn
10 Sontag, E., Nunbhakdi-Craig, V., Sontag, J. M., Diaz-Arrastia, R., Ogris, E., Dayal, S., Lentz, S. R., Arning, E. and Bottiglieri, T. (2007) Protein phosphatase 2A methyltransferase links homocysteine metabolism with tau and amyloid precursor protein regulation. J. Neurosci. 27, 2751-2759   DOI   ScienceOn
11 Azid, A. K. A., Sawa, Y., Ishikawa, T. and Shibata, H. (2006) Purification and characterization of protein phosphatase 2A from petals of tulip Tulipa gesnerina. J. BMB 39, 671-676
12 Oh, M. K., Park, S. J., Kim N. H. and Kim, I. S. (2006) Protein kinase C-delta stimulates haptoglobin secretion. J. BMB 40, 130-134
13 Ahn, J. H., McAvoy, T., Rakhilin, S. V., Nishi, A., Greengard, P. and Nairn, A. C. (2007) Protein kinase A activates protein phosphatase 2A by phosphorylation of the B56delta subunit. Proc. Natl. Acad. Sci. U.S.A. 104, 2979- 2984   DOI   ScienceOn
14 Nishi, A., Bibb, J. A., Snyder, G. L., Higashi, H., Nairn, A. C. and Greengard, P. (2000) Amplification of dopaminergic signaling by a positive feedback loop. Proc. Natl. Acad. Sci. U.S.A. 97, 12840-12845   DOI   ScienceOn
15 Janssens, V. and Goris, J. (2001) Protein phosphatase 2A:a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417-439   DOI   ScienceOn
16 Batut, J., Schmierer, B., Cao, J., Raftery, L. A., Hill, C. S. and Howell, M. (2008) Two highly related regulatory subunits of PP2A exert opposite effects on TGF-beta/Activin/Nodal signalling. Development 135, 2927-2937   DOI   ScienceOn
17 Liu, R., Zhou, X. W., Tanila, H., Bjorkdahl, C., Wang, J. Z., Guan, Z. Z., Cao, Y., Gustafsson, J. A., Winblad, B. and Pei, J. J. (2008) Phosphorylated PP2A (tyrosine 307) is associated with Alzheimer neurofibrillary pathology. J. Cell. Mol. Med. 12, 241-257   DOI   ScienceOn