The Proteolytic Fragments of the Alzheimer's Disease-associated Presenilin-1 Form Heterodimers and Occur as a 100–150-kDa Molecular Mass Complex

Capell, Anja; Grünberg, Jürgen; Pesold, Brigitte; Diehlmann, Anke; Citron, Martin; Nixon, Ralph A.; Beyreuther, Konrad; Selkoe, Dennis J.; Haass, Christian

doi:10.1074/jbc.273.6.3205

Cited by 306 publications

(271 citation statements)

References 44 publications

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“…It is ideal for analysis and partial isolation of membrane Carbonate washed SY5Y membranes (500 lg) were run on BN/PAGE, proteins electroeluted from the PS complex region, treated with reducing sample buffer and analysed on 10% Tris/tricine gels using mini Biorad apparatus. PS1 NTF and full-length PS1 were detected with PS1 [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] antibody, PS1 loop antibody detected PS1 CTF, NCT was detected at 140 kDa, PEN-2 at 10 kDa and PS2 CTF detected with PS2 CTF antibody. Parallel poly(vinylidene difluoride) strips were also probed with APH 1a and 1b antibodies but APH-1 was not detected.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of presenilin complexes from mouse and human brain using Blue Native gel electrophoresis reveals high expression in embryonic brain and minimal change in complex mobility with pathogenic presenilin mutations

Culvenor

Ilaya

Ryan

et al. 2003

European Journal of Biochemistry

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The presenilin proteins are required for intramembrane cleavage of a subset of type 1 membrane proteins including the Alzheimer's disease amyloid precursor protein. Previous studies indicate presenilin proteins form enzymatically active high molecular mass complexes consisting of heterodimers of N‐ and C‐terminal fragments in association with nicastrin, presenilin enhancer‐2 and anterior pharynx defective‐1 proteins. Using Blue Native gel electrophoresis (BN/PAGE) we have studied endogenous presenilin 1 complex mass, stability and association with nicastrin, presenilin enhancer‐2 and anterior pharynx defective‐1. Solubilization of mouse or human brain membranes with dodecyl‐d‐maltoside produced a 360‐kDa species reactive with antibodies to presenilin 1. Presenilin 1 complex levels were high in embryonic brain. Complex integrity was sensitive to Triton X‐100 and SDS, but stable to reducing agent. Addition of 5 m urea caused complex dissolution and nicastrin to migrate as a subcomplex. Nicastrin and presenilin enhancer‐2 were detected in the presenilin 1 complex following BN/PAGE, electroelution and second‐dimension analysis. Anterior pharynx defective‐1 was detected as an 18‐kDa form and 9‐kDa C‐terminal fragment by standard SDS/PAGE of mouse tissues, and as a predominant 36‐kDa band after presenilin 1 complex second‐dimension analysis. Membranes from brain cortex of Alzheimer's disease patients, or from cases with presenilin 1 missense mutations, indicated no change in presenilin 1 complex mobility. Higher molecular mass presenilin 1‐reactive species were detected in brain containing presenilin 1 exon 9 deletion mutation. This abnormality was confirmed using cells transfected with the same presenilin deletion mutation.

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Section: Discussionmentioning

confidence: 99%

“…Glycerol velocity gradient centrifugation and gel filtration of detergent-solubilized membrane extracts have characterized PS complexes of sizes ranging between 150 kDa and two million kDa [5,17,18]. There is limited knowledge about the assembly and role of the PS complex components other than PS.…”

mentioning

confidence: 99%

Characterization of presenilin complexes from mouse and human brain using Blue Native gel electrophoresis reveals high expression in embryonic brain and minimal change in complex mobility with pathogenic presenilin mutations

Culvenor

Ilaya

Ryan

et al. 2003

European Journal of Biochemistry

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“…PSs undergo endoproteolytic cleavage between transmembranes 6 and 7 to generate an N-and Cterminal fragment (NTF and CTF, respectively) (15). The NTF and CTF remain stably associated with each other in a high molecular weight complex (13). Because significant levels of PS NTF and CTF are detected in native neurons while PS holoprotein is virtually undetectable, it is widely assumed that the NTF and CTF are the active components of PS.…”

mentioning

confidence: 99%

“…However, even if PSs are the elusive ␥-secretase, there is some evidence that suggests that they do not act in isolation. For example, these proteins are found in high molecular weight complexes (13,14), their abundance is regulated carefully by an undescribed cellular component (15), and some PS mutations differentially affect A␤ generation and Notch signaling (16).…”

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confidence: 99%

PS1 N- and C-terminal fragments form a complex that functions in APP processing and Notch signaling

Levitan¹,

Lee²,

Song³

et al. 2001

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

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Presenilin proteins play critical roles in the proteolytic processing of both Notch and amyloid precursor protein (APP). Presenilin itself undergoes endoproteolytic processing to generate an N-terminal and C-terminal fragment. As demonstrated previously, overexpression of presenilin 1 (PS1) holoprotein does not change the levels of the N-terminal and C-terminal fragments (NTF and CTF). When we coexpress the PS1 NTF and CTF, marked increases in the cellular levels of these fragments are seen. By coexpressing the PS1 NTF and CTF, we demonstrate conclusively that a noncovalent complex of the NTF and CTF is the active species of presenilin. However, although the PS1 NTF͞CTF complex is necessary for ␥-secretase activity, it is not sufficient. Independent overexpression of the PS1 NTF and CTF was also used to show that the Asp-257 and Asp-385 mutations in PS1 decrease A␤ production by a direct effect on ␥-secretase activity and not by the inhibition of PS1 endoproteolysis.

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“…The residual membrane-bound 'stub' is further processed by an intramembrane (IM) protease, releasing the intracellular domain (ICD), which translocates to the nucleus and modulates gene transcription or other biological activities (Weihofen and Martoglio, 2003). For many of the known eukaryotic RIP substrates, intramembrane proteolysis is catalysed by g-secretase, a multi-protein complex which includes one or both of the presenilin proteases (Capell et al, 1998;Yu et al, 1998;Li et al, 2000;Esler et al, 2002;Takasugi et al, 2002) and a set of associated proteins (Edbauer et al, 2003;Kimberly et al, 2003;Takasugi et al, 2003). Some membrane proteins, such as sterol regulatory element binding protein 2 (SREBP-2) and Notch, appear to signal primarily via the RIP mechanism.…”

Section: Introductionmentioning

confidence: 99%

Regulated proteolysis of the IFNaR2 subunit of the interferon-alpha receptor

et al. 2004

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The type I interferons (IFNs) bind surface receptors, induce JAK kinases and activate STAT transcription factors to stimulate the transcription of genes downstream of IFN-stimulated response elements (ISREs). In this study, we demonstrate that IFNaR2, a subunit of the type I IFN receptor, is proteolytically cleaved in a regulated manner. Immunoblotting shows that multi-step cleavage occurs in response to phorbol ester (PMA) and IFN-a, generating both a transmembrane 'stub' and the intracellular domain (ICD), similar to Notch proteolysis. Isolated membrane fractions process IFNaR2 to release the ICD. A chimeric receptor construct is utilized to show that cleavage requires the presenilins and occurs in response to epidermal growth factor and protein kinase C-d overexpression, as well as PMA and type I IFNs. Fluorescence microscopy demonstrates that a green fluorescent protein-ICD fusion localizes predominantly to the nucleus. A fusion between the ICD and the Gal4 DNA-binding domain represses transcription, in a histone deacetylasedependent manner, of a Gal4 upstream activating sequence-regulated reporter, while overexpression of the ICD alone represses transcription of a reporter linked to an ISRE. Proteolytic cleavage events may facilitate receptor turnover or, more likely, function as a mechanism for signaling similar to that employed by Notch and the Alzheimer's precursor protein.

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The Proteolytic Fragments of the Alzheimer's Disease-associated Presenilin-1 Form Heterodimers and Occur as a 100–150-kDa Molecular Mass Complex

Cited by 306 publications

References 44 publications

Characterization of presenilin complexes from mouse and human brain using Blue Native gel electrophoresis reveals high expression in embryonic brain and minimal change in complex mobility with pathogenic presenilin mutations

Characterization of presenilin complexes from mouse and human brain using Blue Native gel electrophoresis reveals high expression in embryonic brain and minimal change in complex mobility with pathogenic presenilin mutations

PS1 N- and C-terminal fragments form a complex that functions in APP processing and Notch signaling

Regulated proteolysis of the IFNaR2 subunit of the interferon-alpha receptor

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