Endoproteolysis of Presenilin 1 and Accumulation of Processed Derivatives In Vivo

Wang, Shuai; Borchelt, David R.; Lee, Michael K.; Slunt, Hilda H.; Spitzer, Leo; Kim, Grace; Ratovitsky, Tamara; Davenport, Frances; Nordstedt, Christer; Seeger, Mary; Hardy, John; Levey, Aĺlan I.; Gandy, Samuel E.; Jenkins, Nancy A.; Copeland, Neal G.; Price, Donald L.; Sisodia, Sangram S.

doi:10.1016/s0896-6273(00)80291-3

Cited by 983 publications

(1,014 citation statements)

References 32 publications

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“…With hydrophilic loop-specific antiserum B17, in contrast, relatively pronounced 18 -30-kDa fragments were visualized together with intact PS1. These fragments represent most likely carboxyl-terminal fragments of presenilin (35). Remarkably, the observed fragments were not, or only marginally, increased in COS-1 cells overexpressing PS1 (compare lanes 3 and lane 4 in Fig.…”

Section: Immunocytochemical Localization Of Presenilin 1 In the Endopmentioning

confidence: 84%

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Strooper¹,

Beullens²,

Contreras

et al. 1997

Journal of Biological Chemistry

272

187

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Presenilins 1 and 2 are unglycosylated proteins with apparent molecular mass of 45 and 50 kDa, respectively, in transfected COS-1 and Chinese hamster ovary cells. They colocalize with proteins from the endoplasmic reticulum and the Golgi apparatus in transfected and untransfected cells. In COS-1 cells low amounts of intact endogeneous presenilin 1 migrating at 45 kDa are detected together with relative larger amounts of presenilin 1 fragments migrating between 18 and 30 kDa. The presenilins have a strong tendency to form aggregates (mass of 100 -250 kDa) in SDS-polyacrylamide gel electrophoresis, which can be partially resolved when denatured by SDS at 37°C instead of 95°C. Sulfation, glycosaminoglycan modification, or acylation of the presenilins was not observed, but both proteins are posttranslationally phosphorylated on serine residues. The mutations Ala-246 3 Glu or Cys-410 3 Tyr that cause Alzheimer's disease do not interfere with the biosynthesis or phosphorylation of presenilin 1. Finally, using low concentrations of digitonin to selectively permeabilize the cell membrane but not the endoplasmic reticulum membrane, it is demonstrated that the two major hydrophilic domains of presenilin 1 are oriented to the cytoplasm. The current investigation documents the posttranslational modifications and subcellular localization of the presenilins and indicates that postulated interactions with amyloid precursor protein metabolism should occur in the early compartments of the biosynthetic pathway.

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Section: Immunocytochemical Localization Of Presenilin 1 In the Endopmentioning

confidence: 84%

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Strooper¹,

Beullens²,

Contreras

et al. 1997

Journal of Biological Chemistry

272

187

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“…All mutants underwent efficient endoproteolysis (19), as indicated by the lack of full-length PS1 (FL-PS1, 50 kDa), suggesting that neither mutation affected PS1 participation in active γ-secretase assembly (Fig. 3B and C, nonoxidized samples).…”

Section: Disulfide Oxidation Of C92s C410s and C419s Ps1 Mutantsmentioning

confidence: 99%

Deducing the Transmembrane Domain Organization of Presenilin-1 in γ-Secretase by Cysteine Disulfide Cross-Linking

et al. 2006

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Abstractγ-Secretase is a founding member of membrane-embedded aspartyl proteases that cleave substrates within transmembrane domains, and this enzyme is an important target for the development of therapeutics for Alzheimer's disease. The structure of γ-secretase and its precise catalytic mechanism still remain largely unknown. γ-Secretase is a complex of four integral membrane proteins, with presenilin (PS) as the catalytic component. To gain structural and functional information about the 9-transmembrane domain (TMD) presenilin, we employed a cysteine mutagenesis/disulfidecrosslinking approach. Here we report that native Cys92 is close to both Cys410 and Cys419, strongly implying that TMD1 and TMD8 are adjacent to each other. This structural arrangement also suggests that TMD8 is distorted from an ideal helix. Importantly, binding of an active-site directed inhibitor, but not a docking-site directed inhibitor, reduces the ability of the native cysteine pairs of PS1 to crosslink upon oxidation. These findings suggest that the conserved cysteines of TMD1 and TMD8 contribute to or allosterically interact with the active site of γ-secretase.Accumulation of the amyloid β-protein (Aβ) is one of the defining pathological features of Alzheimer's disease. Aβ is produced from the amyloid β-protein precursor (APP) as a result of sequential proteolytic cleavages first by β-secretase (1) and then by γ-secretase (2). γ-Secretase is an aspartyl transmembrane protease that cleaves a number of type I membrane protein substrates, including APP and Notch, in the middle of their transmembrane domains in a poorly understood process of hydrolysis within a hydrophobic environment (3). γ-Secretase is composed of four transmembrane proteins, Aph-1, Pen-2, Nicastrin (NCT) (4-10) and presenilin (PS), which is ostensibly the catalytic component of an unusual aspartyl protease (3,11,12). Despite substantial progress in establishing the full identity of γ-secretase (7-9,13, 14) and the step-wise assembly of the γ-secretase complex (7,15,16), the molecular structure of the protease complex or even any of its individual components remains unknown. PS, as the catalytic component, is of major interest, but only some indirect evidence about its structural arrangement has been reported to date. Indeed, it has only very recently been established that PS1 is a 9-TMD protein (17,18). PS is endoproteolytically processed into an N-terminal fragment (NTF) and C-terminal fragment (CTF) (19). These fragments are metabolically stable, remain associated, and their formation is tightly regulated (20). Also, it is widely † This work was supported by grants to M.S.W. from the National Institutes of Health (NS41355 and AG17574) and the Alzheimer's Association (IIRG-02-4047) and a fellowship to H. L. (Wenner-Gren Foundation in Sweden). An understanding of the mechanism of γ-secretase proteolysis requires a detailed description of the three-dimensional organization of its transmembrane domains. In light of inherent difficulties in obtaining a structure of γ-secr...

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Section: Abnormal Ps1 Endoproteolysis and Notch1 Processingmentioning

confidence: 99%

“…A similar higher 8 molecular weight signal has previously been described in preparations from a patient with a deletion of exon 9 in the PSEN1 gene (PS1-ΔE9) 17 and from SH-SY5Y cells treated with a γ-secretase inhibitor, L-685,458 18 . PS1-ΔE9 and PS1-ΔT440 also cause PS1 endoproteolytic abnormality in a similar manner to the PS1-R278I mutation [19][20][21] .The presence of this high molecular weight γ-secretase may reflect a conformational change in the multi-component complex or binding of additional factor(s) to the complex, although its mechanistic involvement remains unclear.We then examined the effect of the mutation on the metabolism of the γ-secretase substrates. Both the CTF-α and CTF-β of APP and the CTF of N-cadherin accumulated at significant levels in the homozygous PS1-R278I knockin mouse brain but not in the wild-type or heterozygous brains (Fig.…”

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confidence: 99%

Potent amyloidogenicity and pathogenicity of Aβ43

et al. 2011

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Aβ42 is known to be a primary amyloidogenic and pathogenic agent in Alzheimer's disease. However, the role of Aβ43, found just as frequently in patient brains, remains unresolved. We generated knockin mice containing a pathogenic presenilin-1 R278I mutation that causes overproduction of Aβ43. Homozygous mice exhibited embryonic lethality, indicating that the mutation involves loss of function. Crossing amyloid precursor protein transgenic mice with heterozygous mutant mice resulted in elevation of Aβ43 levels, impairment of short-term memory, and acceleration of Aβ pathology, accompanying pronounced accumulation of Aβ43 in plaque cores similar to the biochemical composition observed in patient brains. Consistently, Aβ43 showed a higher propensity to aggregate and was more neurotoxic than Aȕ42. Other pathogenic presenilin mutations also caused overproduction of Aβ43 in a manner correlating with Aβ42 and with age of disease onset. These findings indicate that Aβ43, an overlooked species, is potently amyloidogenic, neurotoxic, and abundant in vivo. 3 Alzheimer's disease, the most common form of dementia, is characterized by two pathological features in the brain, extracellular senile plaques and intracellular neurofibrillary tangles. Senile plaques consist of amyloid-β peptide (Aβ) generated from amyloid precursor protein (APP) through sequential proteolytic processing by β-secretase and γ-secretase. Two major forms of Aβ exist, Aβ40 and Aβ42, with Aβ42 being more neurotoxic due to its higher hydrophobicity, which results in faster oligomerization and aggregation 1 . A number of mutations associated with early-onset familial Alzheimer's disease (FAD) have been identified in the APP, PSEN1 and PSEN2 genes, and these mutations lead to accelerated production of Aβ42 or an increase in the Aβ42/Aβ40 ratio. Together these findings indicate that Aβ42 plays an essential role in the initiation of pathogenesis. However, the possible involvement of longer Aβ species that also exist in Alzheimer's disease brains has not yet been fully investigated.Thus far, various longer Aβ species, such as Aβ43, Aβ45, Aβ48, Aβ49 and Aβ50, have been qualitatively described in Alzheimer's disease brains 2 . Similar Aβ species have also been found in transgenic mice that overexpress APP carrying FAD-linked mutations 3 . Further quantitative studies have revealed that Aβ43 is deposited more frequently than Aβ40 in both sporadic Alzheimer's disease (SAD) and FAD [4][5][6][7] .How these Aβ species with different C-terminal ends are generated from the precursor has mainly been investigated by cell biological and biochemical methods. A number of studies 8,9 demonstrated that γ/ε-cleavage by γ-secretase activity controls the fate of the C-terminal end. Aβ43, generated from Aβ49 via Aβ46, is subsequently converted to Aβ40 by γ-secretase whereas Aβ42 is independently generated from Aβ48 via Aβ45. It has also been reported that the FAD-associated I213T mutation in the PSEN1 gene increases the generation of longer Aβ species, such as Aβ43, Aβ45 a...

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Endoproteolysis of Presenilin 1 and Accumulation of Processed Derivatives In Vivo

Cited by 983 publications

References 32 publications

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Deducing the Transmembrane Domain Organization of Presenilin-1 in γ-Secretase by Cysteine Disulfide Cross-Linking

Potent amyloidogenicity and pathogenicity of Aβ43

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