Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Two transmembrane aspartates in presenilin-1 required for presenilin endoproteolysis and γ-secretase activity

Nature volume 398, pages 513–517 (1999)Cite this article

Abstract

Accumulation of the amyloid-β protein (Aβ) in the cerebral cortex is an early and invariant event in the pathogenesis of Alzheimer's disease. The final step in the generation of Aβ from the β-amyloid precursor protein is an apparently intramembranous proteolysis by the elusive γ-secretase(s)1. The most common cause of familial Alzheimer's disease is mutation of the genes encoding presenilins 1 and 2, which alters γ-secretase activity to increase the production of the highly amyloidogenic Aβ42 isoform2. Moreover, deletion of presenilin-1 in mice greatly reduces γ-secretase activity3, indicating that presenilin-1 mediates most of this proteolytic event. Here we report that mutation of either of two conserved transmembrane (TM) aspartate residues in presenilin-1, Asp 257 (in TM6) and Asp 385 (in TM7), substantially reduces Aβ production and increases the amounts of the carboxy-terminal fragments of β-amyloid precursor protein that are the substrates of γ-secretase. We observed these effects in three different cell lines as well as in cell-free microsomes. Either of the Asp → Ala mutations also prevented the normal endoproteolysis of presenilin-1 in the TM6 → TM7 cytoplasmic loop. In a functional presenilin-1 variant (carrying a deletion in exon 9) that is associated with familial Alzheimer's disease and which does not require this cleavage4, the Asp 385 → Ala mutation still inhibited γ-secretase activity. Our results indicate that the two transmembrane aspartate residues are critical for both presenilin-1 endoproteolysis and γ-secretase activity, and suggest that presenilin 1 is either a unique diaspartyl cofactor for γ-secretase or is itself γ-secretase, an autoactivated intramembranous aspartyl protease.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Expression of wild-type and TM Asp → Ala mutant PS1 holoproteins and CTFs in stable transfectants.
Figure 2: Effect of wild-type and transmembrane Asp → Ala mutant PS1 on APP processing in stable CHO transfectants.
Figure 3: Effect of wild-type and transmembrane Asp → Ala mutant PS1 on APP processing in transient transfectants.
Figure 4: Cell-free generation of Aβ from isolated microsomes, and subcellular distribution of PS1.

Similar content being viewed by others

References

  1. Selkoe, D. J. Cell biology of the amyloid beta-protein precursor and the mechanism of Alzheimer's disease. Annu. Rev. Cell Biol. 10, 373–403 (1994).

    Article  CAS  Google Scholar 

  2. Hardy, J. The Alzheimer family of diseases: many etiologies, one pathogenesis? Proc. Natl Acad. Sci. USA 94, 2095–2097 (1997).

    Article  ADS  CAS  Google Scholar 

  3. De Strooper, B. et al. Deficiency of presenilin-1 inhibits the normal cleavage of amyloid precursor protein. Nature 391, 387–390 (1998).

    Article  ADS  CAS  Google Scholar 

  4. Levitan, D. et al. Assessment of normal and mutant human presenilin function in Caenorhabditis elegans. Proc. Natl Acad. Sci. USA 93, 14940–14944 (1996).

    Article  ADS  CAS  Google Scholar 

  5. Haass, C. et al. Amyloid beta-peptide is produced by cultured cells during normal metabolism. Nature 359, 322–325 (1992).

    Article  ADS  CAS  Google Scholar 

  6. Li, X. & Greenwald, I. Additional evidence for an eight-transmembrane-domain topology for Caenorhabditis elegans and human presenilins. Proc. Natl Acad. Sci. USA 95, 7109–7114 (1998).

    Article  ADS  CAS  Google Scholar 

  7. Thinakaran, G. et al. Endoproteolysis of presenilin 1 and accumulation of processed derivatives invivo. Neuron 17, 181–190 (1996).

    Article  CAS  Google Scholar 

  8. Podlisny, M. B. et al. Presenilin proteins undergo heterogeneous endoproteolysis between Thr291 and Ala299 and occur as stable N- and C-terminal fragments in normal and Alzheimer brain tissue. Neurobiol. Dis. 3, 325–337 (1997).

    Article  CAS  Google Scholar 

  9. Capell, A. et al. The proteolytic fragments of the Alzheimer's disease-associated presenilin-1 form heterodimers and occur as a 100–150-kDa molecular mass complex. J. Biol. Chem. 273, 3205–3211 (1998).

    Article  CAS  Google Scholar 

  10. Venter, J. C., Fraser, C. M., Kerlavage, A. R. & Buck, M. A. Molecular biology of adrenergic and muscarinic cholinergic receptors. Biochem. Pharmacol. 38, 1197–1208 (1989).

    Article  CAS  Google Scholar 

  11. Koo, E. H. & Squazzo, S. L. Evidence that production and release of amyloid beta-protein involves the endocytic pathway. J. Biol. Chem. 269, 17386–17389 (1994).

    CAS  PubMed  Google Scholar 

  12. Xia, W. et al. Enhanced production and oligomerization of the 42-residue amyloid beta-protein by Chinese hamster ovary cells stably expressing mutant presenilins. J. Biol. Chem. 272, 7977–7982 (1997).

    Article  CAS  Google Scholar 

  13. Thinakaran, G. et al. Evidence that levels of presenilins (PS1 and PS2) are coordinately regulated by competition for limiting cellular factors. J. Biol. Chem. 272, 28415–28422 (1997).

    Article  CAS  Google Scholar 

  14. Wolfe, M. S. et al. Asubstrate-based difluoroketone selectively inhibits Alzheimer's gamma-secretase activity. J. Med. Chem. 41, 6–9 (1998).

    Article  CAS  Google Scholar 

  15. Wolfe, M. S. et al. Peptidomimetic probes and molecular modelling suggest that Alzheimer's γ-secretase is an intramembrane-cleaving aspartyl protease. Biochemistry (in the press).

  16. Xia, W. et al. Presenilin 1 regulates the processing of β-amyloid precursor protein C-terminal fragments and the generation of amyloid β-protein in endoplasmic reticulum and Golgi. Biochem. 37, 16465–16471 (1998).

    Article  CAS  Google Scholar 

  17. L'Hernault, S. W. & Arduengo, P. M. Mutation of a putative sperm membrane protein in Caenorhabditis elegans prevents sperm differentiation but not its associated meiotic divisions. J.Cell Biol. 119, 55–68 (1992).

    Article  CAS  Google Scholar 

  18. Li, X. & Greenwald, I. HOP-1, a Caenorhabditis elegans presenilin, appears to be functionally redundant with SEL-12 presenilin and to facilitate LIN-12 and GLP-1 signaling. Proc. Natl. Acad. Sci. USA 94, 12204–12209 (1997).

    Article  ADS  CAS  Google Scholar 

  19. Brockhaus, M. et al. Caspase-mediated cleavage is not required for the activity of presenilins in amyloidogenesis and NOTCH signaling. Neuroreport 9, 1481–1486 (1998).

    Article  CAS  Google Scholar 

  20. Shen, J. et al. Skeletal and CNS defects in presenilin-1-deficient mice. Cell 89, 629–639 (1997).

    Article  CAS  Google Scholar 

  21. Wong, P. C. et al. Presenilin 1 is required for Notch1 and DII1 expression in the paraxial mesoderm. Nature 387, 288–292 (1997).

    Article  ADS  CAS  Google Scholar 

  22. Kovacs, D. M. et al. Alzheimer-associated presenilins 1 and 2: neuronal expression in brain and localization to intracellular membranes in mammalian cells. Nature Med. 2, 224–229 (1996).

    Article  CAS  Google Scholar 

  23. Hua, X., Nohturfft, A., Goldstein, J. L. & Brown, M. S. Sterol resistance in CHO cells traced to point mutation in SREBP cleavage-activating protein. Cell 87, 415–426 (1996).

    Article  CAS  Google Scholar 

  24. Rawson, R. B. et al. Complementation cloning of S2P, a gene encoding a putative metalloprotease required for intramembrane cleavage of SREBPs. Mol. Cell 1, 47–57 (1997).

    Article  CAS  Google Scholar 

  25. Boyartchuk, V. L., Ashby, M. N. & Rine, J. Modulation of Ras and a-factor function by carboxyl-terminal proteolysis. Science 275, 1796–1800 (1997).

    Article  CAS  Google Scholar 

  26. Levitan, D. & Greenwald, I. Effects of SEL-12 presenilin on LIN-12 localization and function in Caenorhabditis elegans. Development 125, 3599–3606 (1998).

    CAS  PubMed  Google Scholar 

  27. Schroeter, R. H., Kisslinger, J. A. & Kopan, R. Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain. Nature 393, 382–385 (1998).

    Article  ADS  CAS  Google Scholar 

  28. De Strooper, B. et al. Apresenilin-1-dependent, γ-secretase-like protease mediates release of Notch intracellular domain. Nature 398, 518–522 (1999).

    Article  ADS  CAS  Google Scholar 

  29. Johnson-Wood, K. et al. Amyloid precursor protein processing and Abeta42 deposition in a transgenic mouse model of Alzheimer disease. Proc. Natl Acad. Sci. USA 94, 1550–1555 (1997).

    Article  ADS  CAS  Google Scholar 

  30. Zhang, J. et al. Subcellular distribution and turnover of presenilins in transfected cells. J. Biol. Chem. 273, 12436–12442 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Watson and J. Zhang for plasmid pCI-C99, and P. Seubert for antibodies 192, 266, 3D6 and 21F12. This work was supported by grants from the NIH to M.S.W. and D.J.S.

Author information

Author notes

  1. Michael S. Wolfe and Weiming Xia: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, 02115, Massachusetts, USA

    Michael S. Wolfe, Weiming Xia, Beth L. Ostaszewski, Thekla S. Diehl, W. Taylor Kimberly & Dennis J. Selkoe

  2. Department of Pharmaceutical Sciences, University of Tennessee, Memphis, 38163, Tennessee, USA

    Michael S. Wolfe

Authors
  1. Michael S. Wolfe
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weiming Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Beth L. Ostaszewski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thekla S. Diehl
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W. Taylor Kimberly
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Dennis J. Selkoe
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Michael S. Wolfe or Dennis J. Selkoe.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wolfe, M., Xia, W., Ostaszewski, B. et al. Two transmembrane aspartates in presenilin-1 required for presenilin endoproteolysis and γ-secretase activity. Nature 398, 513–517 (1999). https://doi.org/10.1038/19077

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/19077

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing