Substrate ectodomain is critical for substrate preference and inhibition of γ-secretase

Funamoto, Satoru; Sasaki, Tōru; Ishihara, Seiko; Nobuhara, Mika; Nakano, Masaki; Watanabe-Takahashi, Miho; Saito, Takashi; Kakuda, Nobuto; Miyasaka, Tomohiro; Nishikawa, Kiyotaka; Saido, Takaomi C.; Ihara, Yasuo

doi:10.1038/ncomms3529

Cited by 53 publications

(50 citation statements)

References 52 publications

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“…Indeed, K m values of ≤1 μM have been reported for APP-, APLP-, ErbB4-, and N-cadheren-based substrates as well as notch (44,56,57). This apparently strong interaction between substrate TMD and γ-secretase is in stark contrast to the binding between rhomboid and its substrate, where no physiologically relevant binding affinity between the two is observed.…”

Section: Discussionmentioning

confidence: 56%

“…We find that γ-secretase preferentially cleaves V1711 over dE notch, which has a longer ectodomain by only 13 amino acids. Similarly, the shorter ectodomain α-secretasegenerated C83 fragment of APP is reported to be a better substrate than the longer β-secretase-generated C99 fragment (44). Like APP, the ectodomains of many other γ-secretase substrates can be cleaved at distinct sites by either α-or β-secretase (12,64,65), resulting in different neo-ectodomain lengths.…”

Section: Discussionmentioning

confidence: 99%

“…Both antibodies and peptides targeting the N terminus of the C99 ectodomain have been found to be effective inhibitors of Aβ production (35,44). Their mode of action was thought to function by blocking the specific nicastrin-substrate ectodomain binding interaction.…”

Section: Discussionmentioning

confidence: 99%

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Nicastrin functions to sterically hinder γ-secretase–substrate interactions driven by substrate transmembrane domain

Bolduc

Montagna

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

129

152

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γ-Secretase is an intramembrane-cleaving protease that processes many type-I integral membrane proteins within the lipid bilayer, an event preceded by shedding of most of the substrate's ectodomain by α-or β-secretases. The mechanism by which γ-secretase selectively recognizes and recruits ectodomain-shed substrates for catalysis remains unclear. In contrast to previous reports that substrate is actively recruited for catalysis when its remaining short ectodomain interacts with the nicastrin component of γ-secretase, we find that substrate ectodomain is entirely dispensable for cleavage. Instead, γ-secretase-substrate binding is driven by an apparent tight-binding interaction derived from substrate transmembrane domain, a mechanism in stark contrast to rhomboid-another family of intramembrane-cleaving proteases. Disruption of the nicastrin fold allows for more efficient cleavage of substrates retaining longer ectodomains, indicating that nicastrin actively excludes larger substrates through steric hindrance, thus serving as a molecular gatekeeper for substrate binding and catalysis.

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Nicastrin functions to sterically hinder γ-secretase–substrate interactions driven by substrate transmembrane domain

Bolduc

Montagna

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

129

152

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“…3, Notch TM -Gal4p cleavage consistently produced 5-fold more ␤-galactosidase than did APP C55 -Gal4p, suggesting that Notch was cleaved at a higher efficiency than that of APP in yeast. It has been shown that ␥-secretase easily captures a short extracellular domain of Notch but has decreased interaction with the long extracellular domain of APP C55 (54). Therefore, a substrate-specific effect of these mutations may be due to substrate preference.…”

Section: Discussionmentioning

confidence: 99%

Suppressor Mutations for Presenilin 1 Familial Alzheimer Disease Mutants Modulate γ-Secretase Activities

Futai

Osawa²,

Cai

et al. 2016

Journal of Biological Chemistry

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␥-Secretase is a multisubunit membrane protein complex containing presenilin (PS1) as a catalytic subunit. Familial Alzheimer disease (FAD) mutations within PS1 were analyzed in yeast cells artificially expressing membrane-bound substrate, amyloid precursor protein, or Notch fused to Gal4 transcriptional activator. The FAD mutations, L166P and G384A (Leu-166 to Pro and Gly-384 to Ala substitution, respectively), were loss-of-function in yeast. We identified five amino acid substitutions that suppress the FAD mutations. The cleavage of amyloid precursor protein or Notch was recovered by the secondary mutations. We also found that secondary mutations alone activated the ␥-secretase activity. FAD mutants with suppressor mutations, L432M or S438P within TMD9 together with a missense mutation in the second or sixth loops, regained ␥-secretase activity when introduced into presenilin null mouse fibroblasts. Notably, the cells with suppressor mutants produced a decreased amount of A␤42, which is responsible for Alzheimer disease. These results indicate that the yeast system is useful to screen for mutations and chemicals that modulate ␥-secretase activity.The ␥-secretase is closely associated with familial Alzheimer disease (FAD) 2 (1). Presenilin is the catalytic subunit of ␥-secretase, which cleaves the intramembrane domain of amyloid precursor protein (APP), thus generating amyloid ␤ (A␤) peptides. More than 150 missense mutations within the two isoforms (PS1 or PS2) of presenilin are identified for the disease. These mutations decrease the generation of A␤ but increase the ratio between A␤42 and A␤40 (2-4), which contain 42 and 40 amino acid residues, respectively. The increased ratio leads to formation of senile plaques in the brains of patients and is responsible for the disease (1). ␥-Secretase modulators also alter the ratios among A␤ species and are classified as A␤42-lowering or -raising compounds (5). The former could represent promising therapeutic agents for the treatment of Alzheimer disease.␥-Secretase, a member of the intramembrane-cleaving protease family, is composed of four membrane proteins as follows: presenilin (PS1 or PS2), nicastrin (NCT), Aph-1 (anterior pharynx-1), and Pen2 (6, 7). Crystal structures of the protease family members, including rhomboid, site-2 protease, and signal peptide peptidases PSH (8 -10), have a membrane-embedded chamber structure, which harbors catalytic residues accessible for water molecules. Although the crystal structure of ␥-secretase is currently unavailable, similar chamber-like structures have been observed by electron microscopy (11,12).Structural analysis, including cysteine-scanning assays, have suggested that PS1 contains nine transmembrane domains (TMD1-9) (13-15), and a hydrophilic "catalytic pore" is formed by TMD1,. The catalytic residues Asp-257 and Asp-385, within TMD6 and -7, respectively, are located in the chamber-like structure (16,17). The catalytic pore resides in the convex side of a horseshoe-like transmembrane arrangement of ␥-secretase, identi...

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“…A␤42 and A␤40 are produced from the cleavage of APP by the ␥-secretase at the Ala713 and Val711 residues, respectively. The previous experiment [15] reported that the production of A␤42 and A␤40 peptides is prevented by binding some peptides to the cleavage site of APP by the ␥-secretase.…”

Section: Introductionmentioning

confidence: 97%

Specific interactions between amyloid-β peptide and curcumin derivatives: Ab initio molecular simulations

Ishimura

Kadoya

Suzuki

et al. 2015

Chemical Physics Letters

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Substrate ectodomain is critical for substrate preference and inhibition of γ-secretase

Cited by 53 publications

References 52 publications

Nicastrin functions to sterically hinder γ-secretase–substrate interactions driven by substrate transmembrane domain

Nicastrin functions to sterically hinder γ-secretase–substrate interactions driven by substrate transmembrane domain

Suppressor Mutations for Presenilin 1 Familial Alzheimer Disease Mutants Modulate γ-Secretase Activities

Specific interactions between amyloid-β peptide and curcumin derivatives: Ab initio molecular simulations

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