From the Center for Neurologic Diseases, Harvard
Medical School and Brigham and Women's Hospital, Boston, Massachusetts
02115 and ¶ Ludwig-Maximilians-University Munich,
Adolf-Butenandt-Institute, Department of Biochemistry, Schillerstrasse
44, D-80336 Muenchen, Germany
It is an aphorism that basic biological research
lays the groundwork for solving important biomedical problems. However,
the reverse is true as well: focusing on problems of human disease can
lead to important new insights into the intricacies of nature. Research
on Alzheimer's disease (AD)1
provides a case study for the latter, where the race to identify molecular players involved in pathogenesis has unveiled some unexpected and fascinating aspects of protease biochemistry and
proteolysis-dependent signal transduction. The
40-42-residue amyloid- Genetic studies of families susceptible to autosomal dominant,
early onset (<60 years) AD underscore the importance of A Despite the loose sequence specificity, The findings that PS mutations affect A FAD-causing presenilin mutants are likewise processed to stable
heterodimers with one exception, a missense mutation in PS1 that leads
to the aberrant splicing out of exon 9, a region that encodes the
endoproteolytic cleavage site (39, 41). This PS1 Given the characterization of The aspartates are critical for The presenilins are also critical for processing of the Notch
receptor, a signaling molecule crucial for cell-fate determination during embryogenesis (57). After translation in the ER, Notch is
processed by a Furin-like protease, resulting in a heterodimeric receptor that is shuttled to the cell surface (Fig. 1A).
Upon interaction with a cognate ligand, the ectodomain of Notch is shed
by a metalloprotease apparently identical to tumor necrosis factor- The parallels between APP and Notch processing are striking. Not only
are both cleaved by TACE, but also the transmembrane regions of both
proteins are processed by a Subcellular localization presents a potential problem for the idea that
presenilins could be Advancing the understanding of More direct evidence that presenilins are the catalytic
components of Taken together, these results strongly suggest that heterodimeric
presenilins contain the catalytic component of The discovery that presenilins are likely As for the origin of presenilins and other intramembrane-cleaving
proteases (I-CLiPs) (85), it seems likely that certain polytopic
integral membrane proteins acquired a few catalytic residues essential
for catalysis rather than that soluble proteases somehow acquired
multiple transmembrane domains. This would explain why polytopic
membrane proteases bear little or no sequence homology with soluble
proteases. For instance, in the S2P family of proteases that process
membrane-bound transcription factors in mammals and bacteria, each
contains an essential HEXXH motif that is a consensus sequence for a number of metalloproteases (86). Otherwise, they bear
little resemblance to other metalloproteases. Similarly, presenilins
require two conserved aspartates for proteolytic function but otherwise
do not look like typical aspartyl proteases. Apparently there are only
a very limited number of efficient biochemical solutions to the problem
of proteolysis, as mechanisms conferred to polytopic proteases are
strikingly similar to those of soluble and membrane-tethered proteases.
Interestingly, the presenilins contain a motif near one of the critical
aspartates that is also found in the bacterial type 4 prepilin protease
family (50, 87), polytopic aspartyl proteases that also have
eight transmembrane domains and require two completely conserved
aspartates (88). This finding further supports the idea that
presenilins may indeed be polytopic aspartyl proteases.
Considerable work lies ahead to understand presenilins/
INTRODUCTION
TOP
INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
peptide (A
) is implicated in the early
pathogenic events that lead to AD, and this peptide is carved out of
the amyloid-
precursor protein (APP) by
- and
-secretases
(Fig. 1A) (1). These two
proteases are considered such important drug targets that the search
for inhibitors of A
production within pharmaceutical companies has gone on for years even though both
- and
-secretases remained unknown until very recently. Last year,
-secretase was identified as
a new membrane-tethered member of the aspartyl protease family (2-6),
a finding that should greatly facilitate drug discovery.
-Secretases, which prevent A
formation by cleaving in the middle of the amyloid domain (7), have now been shown to be members of the
ADAM family (a disintegrin and
metalloprotease) (8, 9). At the same time, the mysterious
-secretase is giving up its secrets as well.
View larger version (55K):
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Fig. 1.
A, proteolytic processing of APP and
Notch. Ectodomain shedding by - or
-secretase generates 83- or
99-residue membrane-associated C-terminal fragments (C83 or C99,
respectively). Metalloproteases such as TACE and ADAM-10 appear to be
-secretases, and
-secretase is a membrane-tethered aspartyl
protease. C83 and C99 are further processed by
-secretase, which
hydrolyzes these substrates in the middle of their transmembrane
regions to form p3 from C83 and A
from C99. Notch is processed by a
Furin-like protease, producing a heterodimeric species that is
trafficked to the cell surface. After ligand binding, the Notch
ectodomain is shed by TACE, and the remaining membrane-associated C
terminus, NEXT, is cut within the transmembrane region by a
-secretase-like protease. B, topology and proteolytic
processing of presenilins. Presenilins possess eight transmembrane
regions and are snipped in the middle of the hydrophobic domain in the
large cytosolic loop between TM 6 and TM 7, producing stable
heterodimers by an unidentified "presenilinase" (PSase).
Two conserved aspartates, one in TM 6 and one in TM 7, are each
required for heterodimer formation and for
-secretase cleavage of
APP and Notch, leading to the hypothesis that presenilins are proteases
activated by autoproteolysis. NTF, N-terminal fragment;
CTF, C-terminal fragment.
Familial Alzheimer's Disease and Generation of Highly
Amyloidogenic A
42
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INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
formation
to the disease process (10, 11). Several different missense mutations
in the APP gene on chromosome 21 cause familial AD (FAD), and these
mutations are found immediately near the
-,
-, or
-secretase
processing sites. A Lys
Asn/Met
Leu double mutation near the
-secretase cleavage site found in a Swedish pedigree leads to
increased A
production (both A
40 and
A
42) (12), and indeed purified
-secretase cuts
peptide substrates with the "Swedish mutation" better than peptides
based on the wild-type sequence (2, 4, 6). The several FAD-causing mutations in APP near the
-secretase processing site also affect A
production, but in these cases the mutations alter the specificity of the protease, leading to increases in the 42-residue A
variant (13, 14). These findings bolstered the "amyloid hypothesis" of AD
pathogenesis, because A
42 is particularly prone to
fibril formation (15) and is the protein initially deposited in the neuritic plaques that characterize the disease (16). Genes encoding the
polytopic presenilin-1 and presenilin-2 (PS1 and PS2), found on
chromosomes 14 and 1, respectively, were later identified as major loci
for FAD (17, 18), and mutations in the presenilins were likewise found
to increase A
42 production in transfected cell lines
(19-21), transgenic animals (19, 20, 22), and human plasma (23). Thus,
presenilins somehow modulate
-secretase activity. In fact, deletion
of PS1 in mice dramatically reduced
-secretase activity (24), and
knockout of both PS1 and PS2 resulted in complete abolition of this APP
processing event (25, 26), demonstrating that presenilins are
absolutely required for proteolysis by
-secretase.
-Secretase carries out an unusual proteolysis within the middle of
the predicted transmembrane domain of its substrates, C99, generated by
-secretase, and C83, generated through alternative processing by
-secretase (Fig. 1A). Indeed, a helical conformation of
the
-secretase cleavage site in APP can explain why nearby FAD-causing mutations specifically increase A
42
production; these mutations are immediately adjacent to the amide bond
processed to give A
42 (27). A phenylalanine-scanning
study near the
-secretase cleavage site and the resulting effects on
A
40 and A
42 production provided important
experimental support for the idea that the substrate is in a helical
conformation upon initial interaction with the protease (28). Other
mutagenesis studies likewise indicated that
-secretase has loose
sequence specificity (29-31).
-secretase nevertheless
appears to process its substrate APP at specific sites. This apparent
paradox can be resolved by considering distance within the membrane as
a key determinant of the site of proteolysis, a concept that together
with the helical model suggests that
-secretase might catalyze an
intramembranous proteolysis. The development of substrate-based
inhibitors led to further indirect characterization of
-secretase;
inhibition by peptidomimetic transition-state analogues suggested an
aspartyl protease mechanism (27). Moreover,
-secretase appears to
have loose sequence specificity for its peptidomimetic inhibitors as it
displays for its substrates (27). Distinctions are observed for the
ability of a given compound to block A
40
vis-à-vis A
42, leading to the
suggestion that these two A
species are generated by different
-secretases (32, 33).
Presenilins and
-Secretases
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INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
42
generation and that PS knockouts prevent A
production raised the
question if presenilins are identical with the
-secretases.
Presenilins appear to bear no obvious sequence homology with known
proteases, and their homology to Spe-4, a Caenorhabditis
elegans protein apparently involved in vesicle transport in
spermatozoa, initially suggested a less direct role in
-secretase
processing of APP (17). Furthermore, overexpression of presenilins does
not lead to increased
-secretase activity (20). However, presenilins
themselves undergo proteolytic processing within the hydrophobic region
of the large cytosolic loop between transmembrane domain (TM) 6 and TM
7 (Fig. 1B) to form stable heterodimeric complexes (39, 40).
(Most studies support an eight-TM topology for the presenilins
(34-36), although six- and seven-TM topologies have also been
suggested (37, 38)). These heterodimers are only produced to limited
levels even upon overexpression of the holoprotein (39, 41-43) and may
be found at the cell surface (44). Expression of exogenous presenilins leads to replacement of endogenous presenilin heterodimers with the
corresponding exogenous heterodimers, indicating competition for
limiting cellular factors needed for stabilization and endoproteolysis (45).
E9 variant is an
active presenilin, like other FAD-causing presenilin mutants, causes
increased A
42 production (19, 46). Upon overexpression, most PS1
E9 is rapidly degraded similar to unprocessed wild-type presenilins; however, a small portion of this PS1 variant is stabilized in cells (41, 47) and forms a high molecular weight complex like the N-
and C-terminal fragments (40, 48), suggesting that it can interact with
the same limiting cellular factors as wild-type presenilins. These
observations are consistent with the idea that the bioactive form of
presenilin is the heterodimer and that the hydrophobic region is an
inhibitory domain. However, several artificial PS variants have been
generated which are biologically active/inactive independent of their
ability to undergo endoproteolysis (49, 50). Therefore, further
evidence is required to finally demonstrate the role of PS
endoproteolysis for their biological and pathological function.
-secretase as an aspartyl
protease that might catalyze an unusual intramembranous proteolysis, the sequences of presenilins were inspected and found to contain two
conserved aspartates predicted to lie within TM 6 and TM 7, flanking
the large cytosolic loop (51). These aspartates appear to align with
each other within the membrane and with the
-secretase cleavage
sites in APP (Fig. 1B). Mutation of either aspartate to
alanine completely prevented PS1 endoproteolysis, and these Asp
Ala
PS1 mutants acted in a dominant-negative manner with respect to
-secretase processing of APP. Similar effects on APP processing were
observed even when conservative mutations to glutamate (51) or
asparagine (49, 52) were made, indicating the crucial identity of these
two residues as aspartates and suggesting that the effects are not
likely because of misfolding. These effects have been corroborated by
several laboratories and have been seen for both PS1 and PS2 (48, 49,
51-54), although a recent study demonstrated that the TM 7 aspartate may be less tolerant of mutagenesis compared with the TM 6 aspartate (55).
-secretase independent of their role
in presenilin endoproteolysis; aspartate mutation in the PS1
E9
variant still blocked
-secretase activity, even though endoproteolysis is not required of this presenilin variant (51). Together these results suggested that presenilins might be the catalytic component of
-secretase; upon interaction with as yet unidentified limiting cellular factors, presenilin undergoes
autoproteolysis via the two aspartates, and the two presenilin subunits
remain together, each contributing one aspartate to the active site of
-secretase. The issue of autoproteolysis, however, needs further investigation, because heterologous expression of a worm PS homologue in human cells leads to a functional PS variant that is not cleaved (56), indicating that a separate "presenilinase" may at least be
required to cleave worm PS.
Presenilins and Notch Processing
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INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
converting enzyme (TACE) (58, 59). Interestingly, metalloproteases such
as TACE and ADAM-10 are among the identified
-secretases that shed
the APP ectodomain (8, 9). The membrane-associated C terminus is then
cut within the postulated transmembrane domain to release the Notch
intracellular domain (NICD), which then translocates to the nucleus
where it interacts with and activates the CSL (CBF1, SU(H), Lag-1)
family of transcription factors (60). NICD formation is absolutely
required for signaling from the Notch receptor; knock-in of a single
point mutation near the transmembrane cleavage site in Notch1 results
in an embryonic lethal phenotype in mice virtually identical to that
observed upon knockout of the entire Notch1 protein (61).
-secretase-like protease that requires
presenilins. Deletion of PS1 in mice is embryonic lethal, with a
phenotype similar to that observed upon knockout of Notch1 (62, 63),
and the PS1/PS2 double knockout phenotype is even more similar (64,
65). Deficiency in PS1 dramatically reduces NICD formation (66), and
the complete absence of presenilins results in total abolition of NICD
production (25, 26). Treatment of cells with
-secretase inhibitors
designed from the transmembrane cleavage site within APP likewise
blocks NICD production (66) and nuclear translocation (67) and reduces
Notch signaling from a reporter gene (67). Moreover, the two conserved
TM aspartates in presenilins are required for cleavage of the Notch TM
domain; as seen with
-secretase inhibitors, expression of Asp mutant PS1 or PS2 results in reduction of NICD formation, translocation, and
signaling (44, 54, 55, 67). Thus, if presenilins are the catalytic
components of the
-secretases that process APP, they are also likely
the catalytic components of the related proteases that clip the
transmembrane region of Notch.
-secretases; presenilins are primarily found in
the ER and early Golgi (68), whereas
-secretase activity apparently
takes place on the cell surface and in the ER and Golgi (69-74). Small
amounts of presenilin heterodimers, however, have been found at the
cell surface in complexation with the membrane-associated C terminus of
Notch (44), and only small amounts of this presumably bioactive form
should be needed for catalysis. Nevertheless, this "spatial
paradox" still needs clear resolution. Another perplexing phenomenon
is that the holoproteins of APP and Notch form stable complexes with
presenilins in the ER (75-77). APP and Notch themselves are not
substrates for
-secretase, so why do they interact with presenilin
if presenilin is the protease? However, as mentioned above, the
membrane-associated C terminus of Notch likewise forms stable complexes
(44), and APP-derived C83 and C99 can also complex with presenilins in
the presence of a
-secretase inhibitor or when either critical
presenilin aspartate is mutated (78). Perhaps presenilins mature
together with APP or Notch. Other unresolved issues are the site and
specificity of Notch cleavage vis à vis that of APP.
Although Notch appears to be cleaved within its transmembrane region,
the site of cleavage is close to the cytosolic edge, whereas the APP
-secretase site is apparently in the middle of the membrane. For
either cleavage event, though, only one of the proteolytic products has
been identified; characterizing the other fragments may clarify this
issue. As for sequence specificity, a single Val to Leu change prevents proteolysis of the Notch transmembrane region (60), whereas a variety
of mutations in the APP transmembrane region is tolerated by
-secretase. Examination of these proteolytic events in purified enzyme assays may offer insights to resolve this problem, especially since recent data suggest that not only APP but also Notch
endoproteolysis occurs largely independent of sequence (79).
Biochemical Evidence That Presenilins Are Proteases
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INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
-secretase and the role of
presenilins in this activity had been hampered by the lack of an isolated enzyme assay. Recently, Li et al. (80) reported a
solubilized
-secretase assay that faithfully reproduces the
properties of the protease activity observed in whole cells. Isolated
microsomes were solubilized with detergent, and
-secretase activity
was determined by measuring A
production from a Flag-tagged version of C99. A
40 and A
42 were produced in the
same ratio as seen in living cells (~9:1), and peptidomimetics that
blocked A
40 and A
42 formation in cells
likewise inhibited production of these A
species in the solubilized
protease assay. After separation of the detergent-solubilized material
by size-exclusion chromatography,
-secretase activity coeluted with
the two subunits of PS1. Remarkably, immunoprecipitated PS1
heterodimers also produced A
from the Flag-tagged substrate,
strongly suggesting that presenilins are part of a large
-secretase complex.
-secretases has recently come from affinity labeling studies using transition-state analogue inhibitors targeted to the
active site of the protease. Shearman et al. (80, 81) identified a peptidomimetic that blocks
-secretase activity with an
IC50 of 0.3 nM in the solubilized protease
assay and contains a hydroxyethyl isostere, a transition-state
mimicking moiety found in many aspartyl protease inhibitors. While the
transition-state mimicking alcohol directs the compound to aspartyl
proteases, flanking substructures determine specificity.
Photoactivatable versions of this compound bound covalently to
presenilin subunits exclusively (82). Interestingly, installation of
the photoreactive group on one end of the inhibitor led to labeling of
the N-terminal presenilin subunit, whereas installation on the other
end resulted in the tagging of the C-terminal subunit. Moreover,
whereas these agents did not label wild-type PS1 holoprotein, they did
tag PS1
E9, which (as described above) is not processed to
heterodimers but nevertheless is active. Similarly, Esler et
al. (83) identified peptidomimetic inhibitors containing a
difluoro alcohol group, another type of transition-state mimicking
moiety, and these compounds were developed starting from a
substrate-based inhibitor designed from the
-secretase cleavage site
in APP. Conversion of one such analogue to a reactive bromoacetamide
provided an affinity reagent that likewise bound covalently and
specifically to PS1 subunits in cell lysates, isolated microsomes, and
whole cells. Either PS1 subunit so labeled could be brought down with
antibodies to the other subunit under coimmunoprecipitation conditions,
demonstrating that the inhibitor bound to heterodimeric PS1. Seiffert
and colleagues (84) likewise identified presenilin subunits as the
molecular target of novel peptidomimetic
-secretase inhibitors.
These inhibitors, however, do not resemble known transition-state
mimics, so it is not clear whether they would be expected to bind to
the active site of the protease.
-secretase; inhibitors in two of the three studies are transition-state analogues targeted to the active site. The active site is likely at the PS
heterodimeric interface; both subunits are labeled by
-secretase affinity reagents, and each contributes one critical aspartate. The
affinity labeling studies also identify the protease responsible for
the transmembrane cleavage of Notch, because substrate-based
-secretase inhibitors like those used by Esler et al.
(83) also block this Notch proteolysis (see above). These
findings reinforce the amyloid hypothesis of AD pathogenesis; all
FAD-causing mutations identified to date (accounting for ~60% of all
FAD cases) are either in the precursor protein leading to A
(APP) or
appear to be located in the proteases that catalyze the final step in A
generation (presenilins/
-secretases). At the same time, an important target for drug development has been identified, although it
is not yet clear whether toxic effects of blocking cleavage of other
-secretase substrates (e.g. Notch) will negate
therapeutic effects. Although presenilins are essential for proper
embryonic development, the role of these proteins in aging adults is
not known.
Perspective
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INTRODUCTION
Familial Alzheimer's Disease...
Presenilins and
-Secretases
Presenilins and Notch...
Biochemical Evidence That...
Perspective
REFERENCES
-secretases raises a
number of fascinating questions with respect to protease biochemistry as well as protease evolution. To carry out hydrolysis in a lipid environment, the transmembrane regions likely form a pore- or channel-like topography, with polar residues facing inward to allow
entry of and interaction with the catalytic water. With such a
sequestered active site and the two-dimensional fluidity of the lipid
bilayer, the substrate probably first interacts with a binding site on
the outer surface of the protease, with subsequent conformational
changes allowing entry of the substrate to the active site within the
protease interior. Further, given that the conformation of the
substrate upon initial interaction with the protease is apparently an
-helix, the enzyme must have some mechanism for unwinding or bending
the substrate near the site of cleavage so that the catalytic water and
protease residues can access the scissile amide bond.
-secretases.
Clearly, the top priority is to identify the other critical components
of the
-secretase complex, allowing reconstitution of activity and
development of a purified enzyme assay. Assembly of the complex into
active
-secretase should allow more focused structural-functional
studies than the current approach of expressing mutant proteins in
whole cells or organisms. Also, understanding how the more than 70 different FAD mutations in presenilins cause specific increases in
A
42 production might shed some light on how this
deleterious function can be skewed in favor of other less noxious A
isoforms. Such understanding should not only suggest novel therapeutic
approaches to AD but should also undergird our appreciation of the
workings of an emerging class of polytopic membrane proteases.
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FOOTNOTES |
---|
* This minireview will be reprinted in the 2001 Minireview Compendium, which will be available in December, 2001.
§ To whom correspondence may be addressed. E-mail: wolfe@cnd.bwh.harvard.edu.
To whom correspondence may be addressed. E-mail:
chaass@pbm.med. uni-muenchen.de.
Published, JBC Papers in Press, December 29, 2000, DOI 10.1074/jbc.R000026200
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ABBREVIATIONS |
---|
The abbreviations used are:
AD, Alzheimer's
disease;
A, amyloid-
peptide;
APP, amyloid-
precursor protein;
FAD, familial AD;
PS, presenilin;
TM, transmembrane domain;
TACE, tumor
necrosis factor-
converting enzyme;
NICD, Notch intracellular
domain;
ER, endoplasmic reticulum.
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REFERENCES |
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1. | Selkoe, D. J. (1999) Nature 399, A23-31[CrossRef][Medline] [Order article via Infotrieve] |
2. |
Vassar, R.,
Bennett, B. D.,
Babu-Khan, S.,
Kahn, S.,
Mendiaz, E. A.,
Denis, P.,
Teplow, D. B.,
Ross, S.,
Amarante, P.,
Loeloff, R.,
et al..
(1999)
Science
286,
735-741 |
3. | Sinha, S., Anderson, J. P., Barbour, R., Basi, G. S., Caccavello, R., Davis, D., Doan, M., Dovey, H. F., Frigon, N., Hong, J., et al.. (1999) Nature 402, 537-540[CrossRef][Medline] [Order article via Infotrieve] |
4. | Yan, R., Bienkowski, M. J., Shuck, M. E., Miao, H., Tory, M. C., Pauley, A. M., Brashier, J. R., Stratman, N. C., Mathews, W. R., Buhl, A. E., et al.. (1999) Nature 402, 533-537[CrossRef][Medline] [Order article via Infotrieve] |
5. | Hussain, I., Powell, D., Howlett, D. R., Tew, D. G., Meek, T. D., Chapman, C., Gloger, I. S., Murphy, K. E., Southa, C. D., Ryan, D. M., et al.. (1999) Mol. Cell. Neurosci. 14, 419-427[CrossRef][Medline] [Order article via Infotrieve] |
6. |
Lin, X.,
Koelsch, G.,
Wu, S.,
Downs, D.,
Dashti, A.,
and Tang, J.
(2000)
Proc. Natl. Acad. Sci. U. S. A.
97,
1456-1460 |
7. | Esch, F. S., Keim, P. S., Beattie, E. C., Blacher, R. W., Culwell, A. R., Oltersdorf, T., McClure, D., and Ward, P. J. (1990) Science 248, 1122-1124[Medline] [Order article via Infotrieve] |
8. |
Lammich, S.,
Kojro, E.,
Postina, R.,
Gilbert, S.,
Pfeiffer, R.,
Jasionowski, M.,
Haass, C.,
and Fahrenholz, F.
(1999)
Proc. Natl. Acad. Sci. U. S. A.
96,
3922-3927 |
9. |
Buxbaum, J. D.,
Liu, K. N.,
Luo, Y.,
Slack, J. L.,
Stocking, K. L.,
Peschon, J. J.,
Johnson, R. S.,
Castner, B. J.,
Cerretti, D. P.,
and Black, R. A.
(1998)
J. Biol. Chem.
273,
27765-27767 |
10. | Hardy, J. (1997) Trends Neurosci. 20, 154-159[CrossRef][Medline] [Order article via Infotrieve] |
11. |
Selkoe, D. J.
(1997)
Science
275,
630-631 |
12. | Citron, M., Oltersdorf, T., Haass, C., McConlogue, L., Hung, A. Y., Seubert, P., Vigo-Pelfrey, C., Lieberburg, I., and Selkoe, D. J. (1992) Nature 360, 672-674[CrossRef][Medline] [Order article via Infotrieve] |
13. | Suzuki, N., Cheung, T. T., Cai, X. D., Odaka, A., Otvos, L., Jr., Eckman, C., Golde, T. E., and Younkin, S. G. (1994) Science 264, 1336-1340[Medline] [Order article via Infotrieve] |
14. |
Tamaoka, A.,
Odaka, A.,
Ishibashi, Y.,
Usami, M.,
Sahara, N.,
Suzuki, N.,
Nukina, N.,
Mizusawa, H.,
Shoji, S.,
Kanazawa, I.,
and Mori, H.
(1994)
J. Biol. Chem.
269,
32721-32724 |
15. | Jarrett, J. T., Berger, E. P., and Lansbury, P. T., Jr. (1993) Biochemistry 32, 4693-4697[Medline] [Order article via Infotrieve] |
16. | Iwatsubo, T., Odaka, A., Suzuki, N., Mizusawa, H., Nukina, N., and Ihara, Y. (1994) Neuron 13, 45-53[Medline] [Order article via Infotrieve] |
17. | Sherrington, R., Rogaev, E. I., Liang, Y., Rogaeva, E. A., Levesque, G., Ikeda, M., Chi, H., Lin, C., Li, G., Holman, K., et al.. (1995) Nature 375, 754-760[CrossRef][Medline] [Order article via Infotrieve] |
18. | Levy-Lahad, E., Wasco, W., Poorkaj, P., Romano, D. M., Oshima, J., Pettingell, W. H., Yu, C. E., Jondro, P. D., Schmidt, S. D., Wang, K., et al.. (1995) Science 269, 973-977[Medline] [Order article via Infotrieve] |
19. | Borchelt, D. R., Thinakaran, G., Eckman, C. B., Lee, M. K., Davenport, F., Ratovitsky, T., Prada, C. M., Kim, G., Seekins, S., Yager, D., et al.. (1996) Neuron 17, 1005-1013[Medline] [Order article via Infotrieve] |
20. | Citron, M., Westaway, D., Xia, W., Carlson, G., Diehl, T., Levesque, G., Johnson-Wood, K., Lee, M., Seubert, P., Davis, A., et al.. (1997) Nat. Med. 3, 67-72[Medline] [Order article via Infotrieve] |
21. |
Tomita, T.,
Maruyama, K.,
Saido, T. C.,
Kume, H.,
Shinozaki, K.,
Tokuhiro, S.,
Capell, A.,
Walter, J.,
Grunberg, J.,
Haass, C.,
Iwatsubo, T.,
and Obata, K.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
2025-2030 |
22. | Duff, K., Eckman, C., Zehr, C., Yu, X., Prada, C. M., Perez-tur, J., Hutton, M., Buee, L., Harigaya, Y., Yager, D., Morgan, D., Gordon, M. N., Holcomb, L., Refolo, L., Zenk, B., Hardy, J., and Younkin, S. (1996) Nature 383, 710-713[CrossRef][Medline] [Order article via Infotrieve] |
23. | Scheuner, D., Eckman, C., Jensen, M., Song, X., Citron, M., Suzuki, N., Bird, T. D., Hardy, J., Hutton, M., Kukull, W., et al.. (1996) Nat. Med. 2, 864-870[Medline] [Order article via Infotrieve] |
24. | De Strooper, B., Saftig, P., Craessaerts, K., Vanderstichele, H., Guhde, G., Annaert, W., Von Figura, K., and Van Leuven, F. (1998) Nature 391, 387-390[CrossRef][Medline] [Order article via Infotrieve] |
25. | Herreman, A., Serneels, L., Annaert, W., Collen, D., Schoonjans, L., and De Strooper, B. (2000) Nat. Cell Biol. 2, 461-462[CrossRef][Medline] [Order article via Infotrieve] |
26. | Zhang, Z., Nadeau, P., Song, W., Donoviel, D., Yuan, M., Bernstein, A., and Yankner, B. A. (2000) Nat. Cell Biol. 2, 463-465[CrossRef][Medline] [Order article via Infotrieve] |
27. | Wolfe, M. S., Xia, W., Moore, C. L., Leatherwood, D. D., Ostaszewski, B., Donkor, I. O., and Selkoe, D. J. (1999) Biochemistry 38, 4720-4727[CrossRef][Medline] [Order article via Infotrieve] |
28. |
Lichtenthaler, S. F.,
Wang, R.,
Grimm, H.,
Uljon, S. N.,
Masters, C. L.,
and Beyreuther, K.
(1999)
Proc. Natl. Acad. Sci. U. S. A.
96,
3053-3058 |
29. |
Tischer, E.,
and Cordell, B.
(1996)
J. Biol. Chem.
271,
21914-21919 |
30. | Maruyama, K., Tomita, T., Shinozaki, K., Kume, H., Asada, H., Saido, T. C., Ishiura, S., Iwatsubo, T., and Obata, K. (1996) Biochem. Biophys. Res. Commun. 227, 730-735[CrossRef][Medline] [Order article via Infotrieve] |
31. | Lichtenthaler, S. F., Ida, N., Multhaup, G., Masters, C. L., and Beyreuther, K. (1997) Biochemistry 36, 15396-15403[CrossRef][Medline] [Order article via Infotrieve] |
32. |
Citron, M.,
Diehl, T. S.,
Gordon, G.,
Biere, A. L.,
Seubert, P.,
and Selkoe, D. J.
(1996)
Proc. Natl. Acad. Sci. U. S. A.
93,
13170-13175 |
33. |
Klafki, H.,
Abramowski, D.,
Swoboda, R.,
Paganetti, P. A.,
and Staufenbiel, M.
(1996)
J. Biol. Chem.
271,
28655-28659 |
34. | Doan, A., Thinakaran, G., Borchelt, D. R., Slunt, H. H., Ratovitsky, T., Podlisny, M., Selkoe, D. J., Seeger, M., Gandy, S. E., Price, D. L., and Sisodia, S. S. (1996) Neuron 17, 1023-1030[Medline] [Order article via Infotrieve] |
35. | Li, X., and Greenwald, I. (1996) Neuron 17, 1015-1021[Medline] [Order article via Infotrieve] |
36. |
Li, X.,
and Greenwald, I.
(1998)
Proc. Natl. Acad. Sci. U. S. A.
95,
7109-7114 |
37. |
Lehmann, S.,
Chiesa, R.,
and Harris, D. A.
(1997)
J. Biol. Chem.
272,
12047-12051 |
38. |
Dewji, N. N.,
and Singer, S. J.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
14025-14030 |
39. | Thinakaran, G., Borchelt, D. R., Lee, M. K., Slunt, H. H., Spitzer, L., Kim, G., Ratovitsky, T., Davenport, F., Nordstedt, C., Seeger, M., et al.. (1996) Neuron 17, 181-190[Medline] [Order article via Infotrieve] |
40. |
Capell, A.,
Grunberg, J.,
Pesold, B.,
Diehlmann, A.,
Citron, M.,
Nixon, R.,
Beyreuther, K.,
Selkoe, D. J.,
and Haass, C.
(1998)
J. Biol. Chem.
273,
3205-3211 |
41. |
Ratovitski, T.,
Slunt, H. H.,
Thinakaran, G.,
Price, D. L.,
Sisodia, S. S.,
and Borchelt, D. R.
(1997)
J. Biol. Chem.
272,
24536-24541 |
42. | Podlisny, M. B., Citron, M., Amarante, P., Sherrington, R., Xia, W., Zhang, J., Diehl, T., Levesque, G., Fraser, P., Haass, C., et al.. (1997) Neurobiol. Dis. 3, 325-337[CrossRef][Medline] [Order article via Infotrieve] |
43. |
Steiner, H.,
Capell, A.,
Pesold, B.,
Citron, M.,
Kloetzel, P. M.,
Selkoe, D. J.,
Romig, H.,
Mendla, K.,
and Haass, C.
(1998)
J. Biol. Chem.
273,
32322-32331 |
44. |
Ray, W. J.,
Yao, M.,
Mumm, J.,
Schroeter, E. H.,
Saftig, P.,
Wolfe, M.,
Selkoe, D. J.,
Kopan, R.,
and Goate, A. M.
(1999)
J. Biol. Chem.
274,
36801-36807 |
45. |
Thinakaran, G.,
Harris, C. L.,
Ratovitski, T.,
Davenport, F.,
Slunt, H. H.,
Price, D. L.,
Borchelt, D. R.,
and Sisodia, S. S.
(1997)
J. Biol. Chem.
272,
28415-28422 |
46. |
Steiner, H.,
Romig, H.,
Grim, M. G.,
Philipp, U.,
Pesold, B.,
Citron, M.,
Baumeister, R.,
and Haass, C.
(1999)
J. Biol. Chem.
274,
7615-7618 |
47. |
Zhang, J.,
Kang, D. E.,
Xia, W.,
Okochi, M.,
Mori, H.,
Selkoe, D. J.,
and Koo, E. H.
(1998)
J. Biol. Chem.
273,
12436-12442 |
48. |
Yu, G.,
Chen, F.,
Nishimura, M.,
Steiner, H.,
Tandon, A.,
Kawarai, T.,
Arawaka, S.,
Supala, A.,
Song, Y. Q.,
Rogaeva, E.,
et al..
(2000)
J. Biol. Chem.
275,
27348-27353 |
49. | Steiner, H., Romig, H., Pesold, B., Philipp, U., Baader, M., Citron, M., Loetscher, H., Jacobsen, H., and Haass, C. (1999) Biochemistry 38, 14600-14605[CrossRef][Medline] [Order article via Infotrieve] |
50. | Steiner, H., Kostka, M., Romig, H., Basset, G., Pesold, B., Hardy, J. A., Capell, A., Meyn, L., Grim, M. G., Baumeister, R., Fechteler, K., and Haass, C. (2000) Nat. Cell Biol. 2, 848-851[CrossRef][Medline] [Order article via Infotrieve] |
51. | Wolfe, M. S., Xia, W., Ostaszewski, B. L., Diehl, T. S., Kimberly, W. T., and Selkoe, D. J. (1999) Nature 398, 513-517[CrossRef][Medline] [Order article via Infotrieve] |
52. | Leimer, U., Lun, K., Romig, H., Walter, J., Grunberg, J., Brand, M., and Haass, C. (1999) Biochemistry 38, 13602-13609[CrossRef][Medline] [Order article via Infotrieve] |
53. |
Kimberly, W. T.,
Xia, W.,
Rahmati, T.,
Wolfe, M. S.,
and Selkoe, D. J.
(2000)
J. Biol. Chem.
275,
3173-3178 |
54. |
Steiner, H.,
Duff, K.,
Capell, A.,
Romig, H.,
Grim, M. G.,
Lincoln, S.,
Hardy, J., Yu, X.,
Picciano, M.,
Fechteler, K.,
et al..
(1999)
J. Biol. Chem.
274,
28669-28673 |
55. | Capell, A., Steiner, H., Romig, H., Keck, S., Baader, M., Grim, M. G., Baumeister, R., and Haass, C. (2000) Nat. Cell Biol. 2, 205-211[CrossRef][Medline] [Order article via Infotrieve] |
56. |
Okochi, M.,
Eimer, S.,
Bottcher, A.,
Baumeister, R.,
Romig, H.,
Walter, J.,
Capell, A.,
Steiner, H.,
and Haass, C.
(2000)
J. Biol. Chem.
275,
40925-40932 |
57. |
Artavanis-Tsakonas, S.,
Rand, M. D.,
and Lake, R. J.
(1999)
Science
284,
770-776 |
58. | Brou, C., Logeat, F., Gupta, N., Bessia, C., LeBail, O., Doedens, J. R., Cumano, A., Roux, P., Black, R. A., and Israël, A. (2000) Mol. Cell 5, 207-216[Medline] [Order article via Infotrieve] |
59. | Mumm, J. S., Schroeter, E. H., Saxena, M. T., Griesemer, A., Tian, X., Pan, D. J., Ray, W. J., and Kopan, R. (2000) Mol. Cell 5, 197-206[Medline] [Order article via Infotrieve] |
60. | Schroeter, E. H., Kisslinger, J. A., and Kopan, R. (1998) Nature 393, 382-386[CrossRef][Medline] [Order article via Infotrieve] |
61. | Huppert, S. S., Le, A., Schroeter, E. H., Mumm, J. S., Saxena, M. T., Milner, L. A., and Kopan, R. (2000) Nature 405, 966-970[CrossRef][Medline] [Order article via Infotrieve] |
62. | Wong, P. C., Zheng, H., Chen, H., Becher, M. W., Sirinathsinghji, D. J., Trumbauer, M. E., Chen, H. Y., Price, D. L., Van der Ploeg, L. H., and Sisodia, S. S. (1997) Nature 387, 288-292[CrossRef][Medline] [Order article via Infotrieve] |
63. | Shen, J., Bronson, R. T., Chen, D. F., Xia, W., Selkoe, D. J., and Tonegawa, S. (1997) Cell 89, 629-639[Medline] [Order article via Infotrieve] |
64. |
Herreman, A.,
Hartmann, D.,
Annaert, W.,
Saftig, P.,
Craessaerts, K.,
Serneels, L.,
Umans, L.,
Schrijvers, V.,
Checler, F.,
Vanderstichele, H.,
et al..
(1999)
Proc. Natl. Acad. Sci. U. S. A.
96,
11872-11877 |
65. |
Donoviel, D. B.,
Hadjantonakis, A. K.,
Ikeda, M.,
Zheng, H.,
Hyslop, P. S.,
and Bernstein, A.
(1999)
Genes Dev.
13,
2801-2810 |
66. | De Strooper, B., Annaert, W., Cupers, P., Saftig, P., Craessaerts, K., Mumm, J. S., Schroeter, E. H., Schrijvers, V., Wolfe, M. S., Ray, W. J., Goate, A., and Kopan, R. (1999) Nature 398, 518-522[CrossRef][Medline] [Order article via Infotrieve] |
67. | Berezovska, O., Jack, C., McLean, P., Aster, J. C., Hicks, C., Xia, W., Wolfe, M. S., Kimberly, W. T., Weinmaster, G., Selkoe, D. J., and Hyman, B. T. (2000) J. Neurochem. 75, 583-593[CrossRef][Medline] [Order article via Infotrieve] |
68. |
Annaert, W. G.,
Levesque, L.,
Craessaerts, K.,
Dierinck, I.,
Snellings, G.,
Westaway, D.,
George-Hyslop, P. S.,
Cordell, B.,
Fraser, P.,
and De Strooper, B.
(1999)
J. Cell Biol.
147,
277-294 |
69. | Haass, C., Koo, E. H., Mellon, A., Hung, A. Y., and Selkoe, D. J. (1992) Nature 357, 500-503[CrossRef][Medline] [Order article via Infotrieve] |
70. | Higaki, J., Quon, D., Zhong, Z., and Cordell, B. (1995) Neuron 14, 651-659[Medline] [Order article via Infotrieve] |
71. | Hartmann, T., Bieger, S. C., Bruhl, B., Tienari, P. J., Ida, N., Allsop, D., Roberts, G. W., Masters, C. L., Dotti, C. G., Unsicker, K., and Beyreuther, K. (1997) Nat. Med. 3, 1016-1020[Medline] [Order article via Infotrieve] |
72. | Cook, D. G., Forman, M. S., Sung, J. C., Leight, S., Kolson, D. L., Iwatsubo, T., Lee, V. M., and Doms, R. W. (1997) Nat. Med. 3, 1021-1023[Medline] [Order article via Infotrieve] |
73. |
Perez, R. G.,
Soriano, S.,
Hayes, J. D.,
Ostaszewski, B.,
Xia, W.,
Selkoe, D. J.,
Chen, X.,
Stokin, G. B.,
and Koo, E. H.
(1999)
J. Biol. Chem.
274,
18851-18856 |
74. |
Wild-Bode, C.,
Yamazaki, T.,
Capell, A.,
Leimer, U.,
Steiner, H.,
Ihara, Y.,
and Haass, C.
(1997)
J. Biol. Chem.
272,
16085-16088 |
75. |
Xia, W.,
Zhang, J.,
Perez, R.,
Koo, E. H.,
and Selkoe, D. J.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
8208-8213 |
76. | Weidemann, A., Paliga, K., Durrwang, U., Czech, C., Evin, G., Masters, C. L., and Beyreuther, K. (1997) Nat. Med. 3, 328-332[Medline] [Order article via Infotrieve] |
77. |
Ray, W. J.,
Yao, M.,
Nowotny, P.,
Mumm, J.,
Zhang, W.,
Wu, J. Y.,
Kopan, R.,
and Goate, A. M.
(1999)
Proc. Natl. Acad. Sci. U. S. A.
96,
3263-3268 |
78. |
Xia, W.,
Ray, W. J.,
Ostaszewski, B. L.,
Rahmati, T.,
Kimberly, W. T.,
Wolfe, M. S.,
Zhang, J.,
Goate, A. M.,
and Selkoe, D. J.
(2000)
Proc. Natl. Acad. Sci. U. S. A.
97,
9299-9304 |
79. | Struhl, G., and Adachi, A. (2000) Mol. Cell 6, 625-636[Medline] [Order article via Infotrieve] |
80. |
Li, Y. M.,
Lai, M. T.,
Xu, M.,
Huang, Q.,
DiMuzio-Mower, J.,
Sardana, M. K.,
Shi, X. P.,
Yin, K. C.,
Shafer, J. A.,
and Gardell, S. J.
(2000)
Proc. Natl. Acad. Sci. U. S. A.
97,
6138-6143 |
81. | Shearman, M. S., Beher, D., Clarke, E. E., Lewis, H. D., Harrison, T., Hunt, P., Nadin, A., Smith, A. L., Stevenson, G., and Castro, J. L. (2000) Biochemistry 39, 8698-8704[CrossRef][Medline] [Order article via Infotrieve] |
82. | Li, Y. M., Xu, M., Lai, M. T., Huang, Q., Castro, J. L., DiMuzio-Mower, J., Harrison, T., Lellis, C., Nadin, A., Neduvelil, J. G., Register, R. B., Sardana, M. K., Shearman, M. S., Smith, A. L., Shi, X. P., Yin, K. C., Shafer, J. A., and Gardell, S. J. (2000) Nature 405, 689-694[CrossRef][Medline] [Order article via Infotrieve] |
83. | Esler, W. P., Kimberly, W. T., Ostaszewski, B. L., Diehl, T. S., Moore, C. L., Tsai, J.-Y., Rahmati, T., Xia, W., Selkoe, D. J., and Wolfe, M. S. (2000) Nat. Cell Biol. 2, 428-434[CrossRef][Medline] [Order article via Infotrieve] |
84. |
Seiffert, D.,
Bradley, J. D.,
Rominger, C. M.,
Rominger, D. H.,
Yang, F.,
Meredith, J. E., Jr.,
Wang, Q.,
Roach, A. H.,
Thompson, L. A.,
Spitz, S. M.,
et al..
(2000)
J. Biol. Chem.
275,
34086-34091 |
85. | Wolfe, M. S., De Los Angeles, J., Miller, D. D., Xia, W., and Selkoe, D. J. (1999) Biochemistry 38, 11223-11230[CrossRef][Medline] [Order article via Infotrieve] |
86. | Rawson, R. B., Zelenski, N. G., Nijhawan, D., Ye, J., Sakai, J., Hasan, M. T., Chang, T. Y., Brown, M. S., and Goldstein, J. L. (1997) Mol. Cell 1, 47-57[Medline] [Order article via Infotrieve] |
87. | Steiner, H., and Haass, C. (2001) Nat. Rev. Mol. Cell Biol., in press |
88. |
LaPointe, C. F.,
and Taylor, R. K.
(2000)
J. Biol. Chem.
275,
1502-1510 |