Familial Alzheimer Disease Presenilin-1 Mutations Alter the Active Site Conformation of γ-secretase

Chau, De-Ming; Crump, Christina J.; Villa, Jennifer C.; Scheinberg, David A.; Li, Yueming

doi:10.1074/jbc.m111.300483

Cited by 58 publications

(78 citation statements)

References 41 publications

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“…Indeed, the qualitative kinetic differences that occur in the profile of A␤ peptides as a result of these familial forms of AD-associated mutations further support the hypothesis that primary amino acid changes in PS, which forms the catalytic active site for ␥-secretase, significantly affect the generation of pathological species of A␤ (37,50). Here, we utilized novel photoaffinity probes and a source of native ␥-secretase from two different cell lines and primary cultures of cortical neurons to document the ability of different GSMs to bind directly to PS1-NTF at different sites.…”

Section: Discussionmentioning

confidence: 69%

“…The pellets were washed once, resuspended in 50 mM MES, 150 mM KCl, 5 mM CaCl 2 , 5 mM MgCl 2 , Complete protease inhibitors (Roche Applied Science), pH ϭ 6, and stored at Ϫ80°C. Membranes from ANP.24 cells overexpressing Aph-1a, nicastrin, and PS1 were prepared as described previously (37). Membranes (800 g) diluted to a volume of 1.2 ml with PBS in 6-or 12-well plates were preincubated with competitor compounds or DMSO control for 30 min at 37°C.…”

Section: Photoaffinity Labeling Of Hela H4-app and Anp24 Membranesmentioning

confidence: 99%

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γ-Secretase Modulator (GSM) Photoaffinity Probes Reveal Distinct Allosteric Binding Sites on Presenilin

Pozdnyakov

Murrey

Crump

et al. 2013

Journal of Biological Chemistry

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116

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Background: Potent GSMs have been identified that lower A␤42; however, the mechanism of modulation is not well understood. Results:The photoaffinity probe E2012-BPyne specifically labels PS1-NTF at a unique site. Conclusion: Acid and imidazole GSMs bind to distinct sites on PS1-NTF and are differentially affected by L458. Significance: Our results provide evidence for multiple binding sites within ␥-secretase that confer specific modulatory effects.

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Section: Discussionmentioning

confidence: 69%

Section: Photoaffinity Labeling Of Hela H4-app and Anp24 Membranesmentioning

confidence: 99%

γ-Secretase Modulator (GSM) Photoaffinity Probes Reveal Distinct Allosteric Binding Sites on Presenilin

Pozdnyakov

Murrey

Crump

et al. 2013

Journal of Biological Chemistry

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116

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“…The majority of AD inherited cases linked to presenilin mutations (185 for PS1 and 13 for PS2) are clustered in the PS1 (ϳ62%) and PS2 (ϳ70%) TMDs. Sequence integrity of each TMD is critical for its integration and folding in the membrane as well as for the structure of the active site pocket (20,21). Recently, the moderate resolution structure of the ␥-secretase complex has been determined by electron cryomicroscopy (22,23).…”

Section: A␤mentioning

confidence: 99%

Presenilin Transmembrane Domain 8 Conserved AXXXAXXXG Motifs Are Required for the Activity of the γ-Secretase Complex

Marinangeli¹,

Tasiaux²,

Opsomer³

et al. 2015

Journal of Biological Chemistry

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Background: Presenilin TMD8 contains a conserved AXXXAXXXG motif, involved in transmembrane protein interactions. Results: Mutation of PS AXXXAXXXG motifs strongly impacts ␥-secretase activity. Conclusion: The geometry and activity of ␥-secretase depend on the integrity of the conserved AXXXAXXXG motifs in TMD8. Significance: The PS AXXXAXXXG motif is a key determinant for understanding the structure, assembly, and function of the ␥-secretase complex.

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“…To detect the less abundant cleavage products of PSH, we used an antibody-based, spectroscopic AlphaLISA assay (13)(14)(15). In this assay, the presence of a specific Aβ peptide brings the donor and acceptor beads to close proximity, triggering the transfer of an oxygen molecule from the donor to the acceptor and consequent detection of emission light from the acceptor ( Fig.…”

Section: Significancementioning

confidence: 99%

Cleavage of amyloid precursor protein by an archaeal presenilin homologue PSH

Dang

Wang

et al. 2015

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

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Aberrant cleavage of amyloid precursor protein (APP) by γ-secretase contributes to the development of Alzheimer's disease. More than 200 disease-derived mutations have been identified in presenilin (the catalytic subunit of γ-secretase), making modulation of γ-secretase activity a potentially attractive therapeutic opportunity. Unfortunately, the technical challenges in dealing with intact γ-secretase have hindered discovery of modulators and demand a convenient substitute approach. Here we report that, similar to γ-secretase, the archaeal presenilin homolog PSH faithfully processes the substrate APP C99 into Aβ42, Aβ40, and Aβ38. The molar ratio of the cleavage products Aβ42 over Aβ40 by PSH is nearly identical to that by γ-secretase. The proteolytic activity of PSH is specifically suppressed by presenilin-specific inhibitors. Known modulators of γ-secretase also modulate PSH similarly in terms of the Aβ42/Aβ40 ratio. Structural analysis reveals association of a known γ-secretase inhibitor with PSH between its two catalytic aspartate residues. These findings identify PSH as a surrogate protease for the screening of agents that may regulate the protease activity and the cleavage preference of γ-secretase.A myloid precursor protein (APP) is initially cleaved by β-secretase in the extracellular space, producing a membrane-tethered, 99-residue carboxyl-terminal fragment known as APP C99 (1). APP C99 then undergoes sequential cleavages by γ-secretase, first at the e sites, yielding Aβ49/Aβ48, and eventually at the γ-sites, generating Aβ42/Aβ40/Aβ38 (2-4) (Fig. 1A). The 230-kDa γ-secretase contains four components: presenilin (PS), Pen-2, Aph-1, and nicastrin (NCT), of which PS1 is the target of most mutations derived from early-onset familial Alzheimer's disease (FAD) patients. Rather than abolishing the protease activity of γ-secretase, these mutations are thought to increase the molar ratio of Aβ42 over Aβ40 (2).Among the cleavage products of γ-secretase, Aβ42 is particularly prone to aggregation, leading to formation of β-amyloid plaque in the brain and presumably causing Alzheimer's disease (3). Other FAD-derived mutations map to APP and PS2, lending support to the causal relationship between formation of β-amyloid plaque and Alzheimer's disease (5). Therapeutic intervention of Alzheimer's disease may directly benefit from in vitro investigation of γ-secretase and discovery of its potential modulators (6). Unfortunately, such effort has been hampered by the difficulty in expression, purification, and manipulation of the complex protease. This problem persists despite recent breakthroughs in structural elucidation of γ-secretase and nicastrin (7,8).The intramembrane aspartate protease PSH from the archaeon Methanoculleus marisnigri JR1 shares 19% sequence identity and 53% sequence similarity with human PS1 (hPS1). The signature motifs for catalysis, ΦYDΦΦ (Φ for a hydrophobic residue) on transmembrane segment 6 (TM6) and ΦGΦGD on TM7, are identical between PSH and hPS1. PSH can be readily overexpressed in Escheri...

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Familial Alzheimer Disease Presenilin-1 Mutations Alter the Active Site Conformation of γ-secretase

Cited by 58 publications

References 41 publications

γ-Secretase Modulator (GSM) Photoaffinity Probes Reveal Distinct Allosteric Binding Sites on Presenilin

γ-Secretase Modulator (GSM) Photoaffinity Probes Reveal Distinct Allosteric Binding Sites on Presenilin

Presenilin Transmembrane Domain 8 Conserved AXXXAXXXG Motifs Are Required for the Activity of the γ-Secretase Complex

Cleavage of amyloid precursor protein by an archaeal presenilin homologue PSH

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