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Transition-state analogue inhibitors of γ-secretase bind directly to presenilin-1

Nature Cell Biology volume 2, pages 428–434 (2000)Cite this article

Abstract

The β-amyloid precursor protein (β-APP), which is involved in the pathogenesis of Alzheimer’s disease, and the Notch receptor, which is responsible for critical signalling events during development, both undergo unusual proteolysis within their transmembrane domains by unknown γ-secretases. Here we show that an affinity reagent designed to interact with the active site of γ-secretase binds directly and specifically to heterodimeric forms of presenilins, polytopic proteins that are mutated in hereditary Alzheimer’s and are known mediators of γ-secretase cleavage of both β-APP and Notch. These results provide evidence that heterodimeric presenilins contain the active site of γ-secretase, and validate presenilins as principal targets for the design of drugs to treat and prevent Alzheimer’s disease.

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Figure 1: Design and synthesis of transition-state-analogue affinity reagents for γ-secretase.
Figure 2: Effects of designed peptidomimetic inhibitors on β-APP processing.
Figure 3: Specific covalent binding of γ-secretase inhibitor BrA–1–Bt to an Mr ~21K protein in cell lysates.
Figure 4: Identification of the affinity-labelled Mr ~21K protein as the C-terminal subunit of PS1.
Figure 5: Specific covalent binding of γ-secretase inhibitor BrA–1–Bt to Mr ~21K and ~31K proteins in isolated microsomes.
Figure 6: γ-secretase affinity reagent binds to the C-terminal subunits of PS1 and PS2.
Figure 7: γ-secretase affinity reagent binds to PS1 NTF–CTF heterodimers.
Figure 8: PS1 subunits are labelled by γ-secretase affinity reagent in living, intact cells.

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Acknowledgements

We thank D. Miller and D. Walsh for helpful discussions, W. Ye for help with cell cultures, J. Shen for providing PS1–/– mouse fibroblasts, S. Gandy for antibody Ab14, C. Haass for antibody 2972 and T. Iwatsubo for antibody PS2L. This work was supported by NIH grants NS37537 (to M.S.W.) and AG12749 (to D.J.S.) and by a Pioneer Award from the Alzheimer’s Association (to D.J.S.).

Correspondence and requests for materials should be addressed to M.S.W.

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Authors and Affiliations

  1. Center for Neurologic Diseases, Harvard Medical School and Brigham and Women’s Hospital, 77 Avenue Louis Pasteur, Boston, 02115, Massachusetts, USA

    William P. Esler, W. Taylor Kimberly, Beth L. Ostaszewski, Thekla S. Diehl, Jui-Yi Tsai, Talat Rahmati, Weiming Xia, Dennis J. Selkoe & Michael S. Wolfe

  2. Department of Pharmaceutical Sciences, University of Tennessee, 847 Monroe Avenue, Memphis, 38163, Tennessee, USA

    Chad L. Moore & Michael S. Wolfe

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  1. William P. Esler
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Correspondence to Michael S. Wolfe.

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Esler, W., Kimberly, W., Ostaszewski, B. et al. Transition-state analogue inhibitors of γ-secretase bind directly to presenilin-1. Nat Cell Biol 2, 428–434 (2000). https://doi.org/10.1038/35017062

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