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An APP inhibitory domain containing the Flemish mutation residue modulates γ-secretase activity for Aβ production

Nature Structural & Molecular Biology volume 17, pages 151–158 (2010)Cite this article

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Abstract

γ-secretase is an aspartyl protease that cleaves multiple substrates within their transmembrane domains. γ-secretase processes the amyloid precursor protein (APP) to generate γ-amyloid (Aγ) peptides associated with Alzheimer's disease. Here, we show that APP possesses a substrate inhibitory domain (ASID) that negatively modulates γ-secretase activity for Aγ production by binding to an allosteric site within the γ-secretase complex. Alteration of this ASID by deletion or mutation, as is seen with the Flemish mutation (A21G), reduces its inhibitory potency and promotes Aγ production. Notably, peptides derived from ASID show selective inhibition of γ-secretase activity for Aγ production over Notch1 processing. Therefore, this mode of regulation represents an unprecedented mechanism for modulating γ-secretase, providing insight into the molecular basis of Alzheimer's disease pathogenesis and a potential strategy for the development of therapeutics.

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Figure 1: The processing of the amyloid precursor protein (APP) by α-, β- and γ-secretases.
Figure 2: Effect of internal Aβ FAD mutations on γ-secretase activity.
Figure 3: Deletion of an ASID generates a more efficient substrate of γ-secretase both in vitro and in cells.
Figure 4: The ASID peptide noncompetitively inhibits γ-secretase.
Figure 5: Photoactivable ASID peptide directly binds to γ-secretase.
Figure 6: Effect of ASID binding on the photoinsertion of photoactivatable transition state inhibitors JC8 and GY-4 into PS1–NTF.
Figure 7: Retro-inverso peptide derivatives of ASID selectively inhibit cellular γ-secretase activity for the processing of APP but not Notch1.

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Acknowledgements

We thank D. Scheinberg, J. Shafer, R. Zhou and S. Gardell for advice and comments on the manuscript, C.C. Shelton, L. Placanica and C. Crump for discussions and suggestions regarding this manuscript, H. Zheng (Baylor College of Medicine) for the APPc antibody, M.-t. Lai (Merck Research Laboratories) for the PS-1 NTF antibody, C.T. Walsh (Harvard Medical School) for the pIAD16 plasmid and R. Kopan for the ΔE-Notch1 construct (Washington University School of Medicine). This work is supported by US National Institutes of Health grant AG026660 (Y.-M.L.), the Alzheimer's Association (Zenith Fellows Award to Y.-M.L.) and the American Health Assistance Foundation (Y.-M.L.), the Mr. William H. Goodwin and Mrs. Alice Goodwin and the Commonwealth Foundation for Cancer Research, the Experimental Therapeutics Center of Memorial Sloan-Kettering Cancer Center and the William Randolph Hearst Fund in Experimental Therapeutics.

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Authors and Affiliations

  1. Molecular Pharmacology and Chemistry Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA

    Yuan Tian, Bhramdeo Bassit, Deming Chau & Yue-Ming Li

  2. Program of Physiology, Biophysics and Systems Biology, Weill Graduate School of Medical Science, Cornell University, New York, New York, USA

    Yuan Tian

  3. Program of Pharmacology, Weill Graduate School of Medical Science, Cornell University, New York, New York, USA

    Deming Chau & Yue-Ming Li

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Y.T. and Y.-M.L designed the experiments and prepared this manuscript; Y.T. and D.C. synthesized and characterized the peptides; Y.T and B.B. made the constructs, purified the proteins and performed in vitro and cell-based assays; Y.T. conducted the kinetic and photoaffinity labeling experiments.

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Correspondence to Yue-Ming Li.

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Tian, Y., Bassit, B., Chau, D. et al. An APP inhibitory domain containing the Flemish mutation residue modulates γ-secretase activity for Aβ production. Nat Struct Mol Biol 17, 151–158 (2010). https://doi.org/10.1038/nsmb.1743

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