Skip to main content

Advertisement

SpringerLink
Log in

Inhibition of Protein Phosphatase 1 Stimulates Secretion of Alzheimer Amyloid Precursor Protein

  • Original Articles
  • Published:
Molecular Medicine Aims and scope Submit manuscript

Abstract

Background

Aberrant metabolism of the Alzheimer amyloid precursor protein (APP) or its amyloidogenic Aβ fragment is thought to be centrally involved in Alzheimer’s disease. Nonamyloidogenic processing of APP involves its cleavage within the Aβ domain by a protease, termed α-secretase, and release of the large extracellular domain, termed APPS. Secretion of APPS can be stimulated by phorbol esters, activators of protein kinase C, with concurrent inhibition of Aβ production. While the role of protein kinases on APP metabolism has been investigated, considerably less effort has been devoted to elucidating the role played by protein phosphatases. Okadaic acid, a protein phosphatase inhibitor, has been shown to stimulate secretion of APPS, but the identity of the phosphatase involved has not been investigated.

Materials and Methods

The secretion of APPS from COS-1 cells was measured in the absence or presence of various doses of serme/threonine-specific phosphatase inhibitors. Quantitation of the derived IC50 values was used to determine the identity of the phosphatase involved in the control of APP secretion.

Results

The availability of protein phosphatase inhibitors with different relative potencies against the different types of serine/threonine-specific protein phosphatase allowed us to examine which of the four known types of protein phosphatase might be involved in the regulation of APP secretion. Both okadaic acid and calyculin A stimulated the secretion of APP from COS-1 cells in a dose-dependent manner. The half-maximal dose for stimulation of APP secretion was approximately 100-fold higher with okadaic acid than with calyculin A.

Conclusions

The nearly 100-fold difference in the observed IC50 values for okadaic acid and calyculin A implicates a type 1 protein phosphatase in the control of APPS production. Protein phosphatase 1 (PP1) is known to be highly expressed in adult mammalian brain, both in neurons and glia. The identification of a specific phosphatase type in the control of APP secretion opens new avenues to the development of rational therapeutic intervention strategies aimed at the prevention and/or treatment of Alzheimer’s Disease.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Glenner GG, Wong CW. (1984) Alzheimer’s disease: Initial report of the purification and characterization of a novel cerebrovascular amyloid protein. Biochem. Biophys. Res. Commun. 120: 885–890.

    Article  CAS  PubMed  Google Scholar 

  2. Caporaso GL, Gandy SE, Buxbaum JD, Ramabhadran TV, Greengard P. (1992) Protein phosphorylation regulates secretion of Alzheimer β/A4 amyloid precursor protein. Proc. Natl. Acad. Sci. U.S.A. 89: 3055–3059.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Gillespie SL, Golde TE, Younkin SG. (1992) Secretory processing of the Alzheimer amyloid β/A4 protein precursor is increased by protein phosphorylation. Biochem. Biophys. Res. Commun. 187: 1285–1290.

    Article  CAS  PubMed  Google Scholar 

  4. da Cruz e Silva OAB, Iverfeldt K, Oltersdorf T, et al. (1993) Regulated cleavage of Alzheimer β-amyloid precursor protein in the absence of the cytoplasmic tail. Neuroscience 57: 873–877.

    Article  PubMed  Google Scholar 

  5. Hung AY, Selkoe DJ. (1994) Selective ectodomain phosphorylation and regulated cleavage of β-amyloid precursor protein. E.M.B.O. J. 13: 534–542.

    CAS  Google Scholar 

  6. Gandy SE, Czernik AJ, Greengard P. (1988) Phosphorylation of Alzheimer disease amyloid precursor protein peptide by protein kinase C and Ca2+/calmodulin-dependent protein kinase II. Proc. Natl. Acad. Sci. U.S.A. 85: 6218–6221.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Suzuki T, Nairn AC, Gandy SE, Greengard P. (1992) Phosphorylation of Alzheimer amyloid precursor protein by protein kinase C. Neuroscience 48: 755–761.

    Article  CAS  PubMed  Google Scholar 

  8. Shenolikar S, Nairn AC. (1991) Protein phosphatases: Recent progress. Adv. Sec. Mess. Phosphoprot. Res. 23: 1–123.

    CAS  Google Scholar 

  9. da Cruz e Silva OB, da Cruz e Silva EF, Cohen PTW. (1988) Identification of a novel protein phosphatase catalytic subunit by cDNA cloning. F.E.B.S. Lett. 242: 106–110.

    Article  Google Scholar 

  10. Ishihara H, Martin BL, Brautigan DL, et al. (1989) Calyculin A and okadaic acid: Inhibitors of protein phosphatase activity. Biochem. Biophys. Res. Commun. 159: 871–877.

    Article  CAS  PubMed  Google Scholar 

  11. da Cruz e Silva OB, da Cruz e Silva EF, Greengard P. (1994) Role of protein phosphatase 1 in the metabolism of Alzheimer amyloid precursor protein. Soc. Neurosci. Abs. 20: 440.

    Google Scholar 

  12. Weidemann A, Konig G, Bunke D, et al. (1989) Identification, biogenesis and localization of precursors of Alzheimer’s disease A4 amyloid protein. Cell 57: 115–126.

    Article  CAS  PubMed  Google Scholar 

  13. Smith PK, Krohn RI, Hermanson GT, et al. (1985) Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76–85.

    Article  CAS  PubMed  Google Scholar 

  14. Laemmli UK. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  15. Buxbaum JD, Gandy SE, Cicchetti P, et al. (1990) Processing of Alzheimer β/A4 amyloid precursor protein: Modulation by agents that regulate protein phosphorylation. Proc. Natl. Acad. Sci. U.S.A. 87: 6003–6006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wasco W, Bupp K, Magendantz M, Gusella JF, Tanzi RE, Solomon F. (1992) Identification of a mouse brain cDNA that encodes a protein related to the Alzheimer disease-associated amyloid beta protein precursor. Proc. Natl. Acad. Sci. U.S.A. 89: 10758–10762.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Wasco W, Gurubhagavatula S, Paradis MD, et al. (1993) Isolation and characterization of APLP2 encoding a homologue of the Alzheimer’s associated amyloid beta protein precursor. Nature Genet. 5: 95–100.

    Article  CAS  PubMed  Google Scholar 

  18. Li YM, Casida JE. (1992) Cantharidin-binding protein: Identification as protein phosphatase 2A. Proc. Natl. Acad. Sci. U.S.A. 89: 11867–11870.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Li YM, Mackintosh C, Casida JE. (1993) Protein phosphatase 2A and its [3H] cantharidin/[3H] endothall thioanhydride binding site. Inhibitor specificity of cantharidin and ATP analogues. Biochem. Pharmacol. 46: 1435–1443.

    Article  CAS  PubMed  Google Scholar 

  20. Sasaki K, Shima H, Kitagawa Y, Irino S, Sugimura T, Nagao M. (1990) Identification of members of the protein phosphatase 1 gene family in the rat and enhanced expression of protein phosphatase 1a gene in rat hepatocellular carcinomas. Jpn. J. Cancer Res. 81: 1272–1280.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. da Cruz e Silva EF, Greengard P. (1995) Cloning of neuronal isoforms of protein phosphatase 1 by low-stringency screening of cDNA libraries. Neuroprotocols 6: 2–10.

    Google Scholar 

  22. da Cruz e Silva EF, Fox CA, Ouimet CC, Gustafson E, Watson SJ, Greengard P. (1995) Differential expression of protein phosphatase 1 isoforms in mammalian brain. J. Neuroscience 15: 3375–3389.

    Article  Google Scholar 

  23. Ouimet CC, da Cruz e Silva EF, Greengard P. (1995) The α and γ1 isoforms of protein phosphatase 1 are highly and specifically concentrated in dendritic spines. Proc. Natl. Acad. Sci. U.S.A. 92: 3396–3400.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Hescheld J, Mieskes G, Ruegg JC, Takai A, Trautwein W. (1988) Effects of a protein phosphatase inhibitor, okadaic acid, on membrane currents of isolated guinea-pig cardiac myocytes. Pfluegers Arch. 412: 248–252.

    Article  Google Scholar 

  25. Bialojan C, Takai A. (1988) Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics. Biochem J. 256: 283–290.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Starke LC, Jennings ML. (1993) K-Cl co-transport in rabbit red cells: Further evidence for regulation by protein phosphatase type 1. Am. J. Physiol. 264: C118–C124.

    Article  CAS  PubMed  Google Scholar 

  27. Murata K, Sakon M, Kambayashi J, et al. (1993) The possible involvement of protein phosphatase 1 in thrombin-induced Ca2+ influx of human platelets. J. Cell. Biochem. 51: 442–445.

    Article  CAS  PubMed  Google Scholar 

  28. Walaas SI, Aswad DW, Greengard P. (1983) A dopamine- and cyclic AMP-regulated phosphoprotein enriched in dopamine-innervated brain regions. Nature 301: 69–71.

    Article  CAS  PubMed  Google Scholar 

  29. Snyder GL, Girault J-A, Chen JY, et al. (1992) Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: Regulation by factors other than dopamine. J. Neurosci. 12: 3071–3083.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Snyder G, Fisone G, Greengard P. (1994) Phosphorylation of DARPP-32 is regulated by GABA in rat striatum and substantia nigra. J. Neurochem. 63: 1766–1771.

    Article  CAS  PubMed  Google Scholar 

  31. Hemmings Jr HC, Greengard P, Tung HYL, Cohen P. (1984) DARPP-32, a dopamine-regulated neuronal phosphoprotein, is a potent inhibitor of protein phosphatase-1. Nature 310: 503–505.

    Article  CAS  PubMed  Google Scholar 

  32. Halpain S, Girault J-A, Greengard P. (1990) Activation of NMDA receptors induces dephosphorylation of DARPP-32 in rat striatal slices. Nature 343: 369–372.

    Article  CAS  PubMed  Google Scholar 

  33. Davies P, Maloney AJF. (1976) Selective loss of central cholinergic neurons in Alzheimer’s disease. Lancet 2: 1403.

    Article  CAS  PubMed  Google Scholar 

  34. Tomlinson BE, Corsellis JAN. (1984) Ageing and the dementias. In: Adams JH, Corsellis JAN, Duchen LW (eds). Greenfield’s Neuropathology. 4th Ed. John Wiley & Sons, New York, pp. 951–1025.

    Google Scholar 

  35. Cole G, Dobkins KR, Hansen LA, Terry RD, Saitoh T. (1988) Decreased levels of protein kinase C in Alzheimer brain. Brain Res. 452: 165–174.

    Article  CAS  PubMed  Google Scholar 

  36. Van Huynh T, Cole G, Katzman R, Huang K-P, Saitoh T. (1989) Reduced protein kinase C immunoreactivity and altered protein phosphorylation in Alzheimer’s disease fibroblasts. Arch. Neurol. 46: 1195–1199.

    Article  PubMed  Google Scholar 

  37. Masliah E, Cole GM, Hansen LA, et al. (1991) Protein kinase C alteration is an early biochemical marker in Alzheimer’s disease. J. Neurosci. 11: 2759–2767.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Buxbaum JD, Oishi M, Chen HI, et al. (1992) Cholinergic agonists and interleukin 1 regulate processing and secretion of the Alzheimer β/A4 amyloid precursor protein. Proc. Natl. Acad. Sci. U.S.A. 89: 10075–10078.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Nitsch RM, Slack BE, Wurtman RJ, Growdon JH. (1992) Release of Alzheimer amyloid precursor derivatives stimulated by activation of muscarinic acetylcholine receptors. Science 258: 304–307.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Drs. J. D. Buxbaum, S. E. Gandy, and A. C. Nairn for critical reading of the manuscript. This work was supported by U. S. Public Health Service Grant AG-09464. CTBVZ was supported by a postdoctoral fellowship from CNPq and by PICD from Universidade Estadual de Londrina (Brazil).

Author information

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, Box 296, 1230 York Avenue, New York, New York, 10021, USA

    Edgar F. da Cruz e Silva, Odete A. B. da Cruz e Silva, Cássia Thais B. V. Zaia & Paul Greengard

Authors
  1. Edgar F. da Cruz e Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Odete A. B. da Cruz e Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Cássia Thais B. V. Zaia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paul Greengard
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

da Cruz e Silva, E.F., da Cruz e Silva, O.A.B., Zaia, C.T.B.V. et al. Inhibition of Protein Phosphatase 1 Stimulates Secretion of Alzheimer Amyloid Precursor Protein. Mol Med 1, 535–541 (1995). https://doi.org/10.1007/BF03401590

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03401590

Search

Navigation