COMMUNICATION
The Biological and Pathological Function of the Presenilin-1
Exon 9 Mutation Is Independent of Its Defect to Undergo Proteolytic
Processing*
Harald
Steiner
,
Helmut
Romig§,
Melissa G.
Grim¶,
Uwe
Philipp§,
Brigitte
Pesold
,
Martin
Citron
,
Ralf
Baumeister¶**, and
Christian
Haass
**
From the
Central Institute of Mental Health,
Department of Molecular Biology, J5, 68159 Mannheim, Germany,
§ Boehringer Ingelheim KG, CNS Research, 55216 Ingelheim,
Germany, ¶ Genzentrum, Feodor-Lynen-Str.25, 81377 Munich, Germany,
and
Amgen Inc., Thousand Oaks, California 91320-1789
 |
ABSTRACT |
The two homologous presenilins are
key factors for the generation of amyloid
-peptide (A
), since
Alzheimer's disease (AD)-associated mutations enhance the production
of the pathologically relevant 42-amino acid A
(A
42), and a gene
knockout of presenilin-1 (PS1) significantly inhibits total A
production. Presenilins undergo proteolytic processing within the
domain encoded by exon 9, a process that may be closely related to
their biological and pathological activity. An AD-associated mutation
within the PS1 gene deletes exon 9 (PS1
exon9) due to a splicing
error and results in the accumulation of the uncleaved full-length
protein. We now demonstrate the unexpected finding that the
pathological activity of PS1
exon9 is independent of its lack to
undergo proteolytic processing, but is rather due to a point mutation
(S290C) occurring at the aberrant exon 8/10 splice junction.
Mutagenizing the cysteine residue at position 290 to the original
serine residue completely inhibits the pathological activity in regard
to the elevated production of A
42. Like PS1
exon9, the resulting
presenilin variant (PS1
exon9 C290S) accumulates as an uncleaved
protein and fully replaces endogenous presenilin fragments. Moreover,
PS1
exon9 C290S exhibits a significantly increased biological
activity in a highly sensitive in vivo assay as compared
with the AD-associated mutation. Therefore not only the increased
A
42 production but also the decreased biological function of
PS1
exon9 is due to a point mutation and independent of the lack of
proteolytic processing.
 |
INTRODUCTION |
Early onset Alzheimer's disease
(AD)1 can occur due to
mutations within the genes encoding the
-amyloid precursor protein (
APP) and the two presenilins, PS1 and PS2 (Refs. 1-3; summarized in Refs. 4 and 5). The mutations increase the generation of the
42-amino acid version of A
(A
42), which aggregates more readily
(6) and is therefore preferentially deposited in senile plaques (5). PS
proteins are also involved in the physiological production of A
,
since the lack of PS1 expression in PS1
/
mice results
in a dramatically reduced A
production (7). All but one PS mutations
are point mutations affecting conserved amino acids (4, 5). However,
due to a splicing error, the PS1
exon9 mutation results in the
deletion of the domain encoded by exon 9 (8). This domain contains the
cleavage site for proteolytic processing (9, 10), and therefore
PS1
exon9 accumulates as an uncleaved protein (9). Since proteolytic
processing is highly regulated (9, 11, 12) and appears to be altered by
PS mutations (Refs. 13-15; summarized in Ref. 16), the lack of
proteolytic processing caused by the exon 9 deletion was expected to be
responsible for its pathological activity. We now demonstrate the
unexpected finding that the pathological function of PS1
exon9 as
well as its reduced biological activity is independent of its lack to undergo proteolytic processing, but is rather due to a point mutation (S290C) that is the result of the aberrant exon 8/10 splice junction.
 |
EXPERIMENTAL PROCEDURES |
Cell Culture and Cell Lines--
K293 cells were cultured in
Dulbecco's modified Eagle's medium supplemented with 10% fetal
bovine serum, 1% penicillin/streptomycin, 200 µg/ml G418 (to select
for
APP expression) and 200 µg/ml zeocin (to select for presenilin
expression). K293 cells stably expressing PS1
exon9 C290S were
generated by transfection of K293 cells stably expressing
APP
containing the Swedish mutation (17). K293 cells stably transfected
with Swedish
APP695, wt PS1, and PS1
exon9 were described
previously (18).
Construction of the cDNA Encoding PS1
Exon9
C290S--
The cDNA encoding PS1
exon9 C290S was constructed by
mutagenizing the cysteine at codon 290 of the PS1
exon9 cDNA (8)
to serine in a two-step PCR procedure. The following primers were designed: first PCR, PS1-187-F (5'-CCGAATTCAAGAAAGAACCTCAA-3') and
E9-C290S-R (5'-GTGACTCCCTTTCTGTGGAGGAGTAAATGAGAGC-3'); second PCR,
E9-C290S-F (5'-GCTCTCATTTACTCCTCCACAGAAAGGGAGTCAC-3') and PS1-STOP-R (5'-CGCCTCGAGGCAAATATGCTAGATATA-3').
After gel purification the PCR products were mixed and subjected to a
final PCR with primers PS1-187-F and PS1-STOP-R. The resulting PCR
product was digested with EcoRI/XhoI and cloned into the pcDNA3.1/Zeo(+) expression vector (Invitrogen). The
cDNA was sequenced to verify successful mutagenesis.
Antibodies--
The polyclonal antibodies against amino acids
263-407 of PS1 (3027) and amino acids 297-356 of PS2 (3711) were
described previously (19, 20). The monoclonal antibodies to the PS1 and
PS2 loop were raised against fusion proteins containing amino acids
263-407 (BI.3D7) or 297-356 (BI.HF5C).
Metabolic Labeling and Immunoprecipitation of PS--
Stably
transfected K293 cell lines were grown to confluence in 10-cm dishes.
After starvation for 1 h in 4 ml of methionine- and serum-free MEM
(MEM supplemented with 1% L-glutamine and 1% penicillin/streptomycin) cells were metabolically labeled with 700 µCi of [35S]methionine (Promix, Amersham Pharmacia
Biotech) in 4 ml of methionine- and serum-free MEM for 1 h. Cell
extracts were prepared and subjected to immunoprecipitation of PS as
described (12). PS immunoprecipitates were solubilized in sample buffer
containing 4 M urea for 10 min at 65 °C, separated on
SDS-urea gels, and analyzed by fluorography (19).
Combined Immunoprecipitation/Western Blotting--
Stably
transfected K293 cell lines were grown to confluence. Cell extracts
were prepared and subjected to immunoprecipitation as described (12).
Following gel electrophoresis, immunoprecipitated proteins were
identified by immunoblotting (12). Bound antibodies were detected by
enhanced chemiluminescence (ECL, Amersham Pharmacia Biotech).
Analysis of A
40 and A
42--
Culture medium (2 ml) was
collected from confluent K293 cells grown in six-well dishes for
24 h. The medium was assayed for A
40 and A
42 using a
previously described ELISA assay (12). A
peptides were
immunoprecipitated using antibody 6E10 (Senetek) according to
established protocols (21).
Transgenic Lines of Caenorhabditis elegans and Rescue
Assays--
Expression constructs were generated as described
previously (22). Transgenic lines were established by microinjection of plasmid DNA mixtures into the C. elegans germ line to create
extrachromosomal arrays as described previously (22). All plasmids used
in this study were injected at a concentration of 20 ng/µl into
sel-12(ar171) or sel-12(ar171)
unc-1(e538) hermaphrodites along with ttx-3:GFP as a cotransformation marker. Successful transformation was monitored by the expression of GFP in the AIY interneurons of F1 and F2 generation animals. Four independent lines from the progeny of F2
generation animals were established.
As the sel-12(ar171) animals never lay eggs (23), rescue of
the sel-12 defect can be quantified by scoring egg laying
behavior in transgenic animals (22, 24). Consequently, for each
transgenic line, we examined 50 transgenic animals for their ability to
lay eggs and determined their brood size. The number of eggs laid by
individual transgenic animals was counted and placed into four categories: "Egl+++," robust egg laying, more than 30 eggs laid (wt
phenotype); "Egl++," 15-30 eggs laid; "Egl+," 5-15 eggs laid; "Egl
," no eggs laid.
 |
RESULTS |
The PS1
exon9 mutation changes a G to a T at the splice site for
exon 9. It therefore destroys the minimal required consensus sequence
for the splice acceptor site (8), which then results in an aberrant
deletion of the domain encoded by exon 9 (Ref. 8; Fig.
1A). However, the mutation
also changes codon 290 at the exon 8/10 splice junction from a serine
to a cysteine (Ref. 8; Fig. 1A). We therefore have
investigated if the single amino acid exchange or the deletion of the
domain required for proteolytic processing is responsible for the
pathological activity of the PS1
exon9 mutation in regard to A
42
generation. For this purpose we mutagenized amino acid 290 of
PS1
exon9 to a serine (PS1
exon9 C290S), thus correcting the point
mutation (Fig. 1A). cDNAs encoding PS1
exon9 and
PS1
exon9 C290S were stably transfected into kidney 293 cells
overexpressing
APP containing the Swedish mutation. This cell line
was used previously to determine the pathological effect of PS
mutations on A
production (12, 18). Untransfected K293 cells
expressing endogenous PS, as well as cell lines overexpressing PS1
exon9 and PS1
exon9 C290S were pulse-labeled with
[35S]methionine for 1 h, and cell lysates were
immunoprecipitated with antibody 3027 to the large loop of PS1. Both
PS1
exon9 and PS1
exon9 C290S accumulated as full-length proteins
(Fig. 1B). As reported previously (9, 10) very little
endogenous full-length PS1 could be detected in the untransfected cell
line.

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Fig. 1.
Generation and expression of
PS1 exon9 and PS1 exon9
C290S. A, schematic representation of the genomic
structure (exon 8 to exon 10) of wt PS1, PS1 exon9 and the amino acid
sequence at the splice junctions of exons 8, 9, and 10. The nucleotide
exchange (G to T) in the PS1 gene identified in the DNA of patients
with the PS1 exon9-linked mutation results in an amino acid exchange
at the exon 8/10 splice junction, which changes codon 290 from a serine
to a cysteine (bold letters). B, K293 cells
expressing endogenous presenilins (control) as well as K293 cells
stably overexpressing PS1 exon9, or PS1 exon9 C290S were pulse
labeled with [35S]methionine for 1 h, and cell
lysates were immunoprecipitated with antibody 3027 (19) to the large
loop of PS1. Full-length PS (fl-PS) proteins accumulate in
the cell lines expressing PS1 exon9 and PS1 exon9 C290S but not in
cells expressing endogenous presenilins. The asterisk
indicates dimeric forms of PS1 (19). C and
D, stable expression of PS1 exon9 and PS1 exon9
C290S inhibits formation of endogenous PS1 (C) and PS2 C-terminal
fragments (CTF) (D). Cell lysates from K293 cells
and cell lines overexpressing PS1 exon9 and PS1 exon9 C290S were
immunoprecipitated with the previously described antibodies (see
"Experimental Procedures") specific to the large loop of PS1 (3027;
Ref. 19) and PS2 (3711; Ref. 20) and detected by immunoblotting using
the monoclonal antibodies BI.3D7 and BI.HF5C.
|
|
It has been shown before that overexpression of PS proteins results in
the replacement of endogenous PS fragments and the accumulation of
PS1
exon9 prevents the formation of stable PS fragments (9, 11, 12).
To prove that overexpression of PS1
exon9 C290S still allows the
reduction of endogenous PS fragments, cell lysates from unlabeled cells
were immunoprecipitated with antibodies specific to the PS1 or PS2 loop
domain (see "Experimental Procedures"), and PS fragments were
identified with the PS1/PS2-specific monoclonal antibodies BI.3D7 and
BI.HF5C (see "Experimental Procedures"). Whereas cell lines
expressing endogenous presenilins produced PS1 as well as PS2
C-terminal fragments (Fig. 1, C and D),
overexpression of PS1
exon9 and PS1
exon9 C290S inhibited the
formation of endogenous PS1 and PS2 fragments (Fig. 1, C and
D). This demonstrates that PS1
exon9 C290S, like
PS1
exon9, fully replaced presenilins derived from the endogenous
genes and also proves that PS1
exon9 C290S does not undergo
proteolytic processing but rather accumulates as an uncleaved
full-length protein.
In order to investigate the pathological activity of the mutant PS
derivatives on A
production, control cells as well as cells
overexpressing PS1
exon9 and PS1
exon9 C290S were metabolically labeled with [35S]methionine, and A
40 and A
42
peptides were immunoprecipitated from the conditioned medium using
antibody 6E10. This antibody is raised to A
1-17 and therefore
recognizes A
40 as well as A
42 (see "Experimental
Procedures"). Isolated A
peptides were then separated on a
previously described gel system, which allows the specific resolution
of A
40 and A
42 (25). As shown in Fig. 2A, cells overexpressing PS1
exon9
secreted elevated levels of A
42 as compared with control cells. In
contrast, the cell line stably transfected with PS1
exon9 C290S
produced significantly lower amounts of A
42 as cells expressing the
Alzheimer's disease-associated PS1
exon9 mutation. This suggests
that the pathological activity of the PS1
exon9 mutation is due to
the point mutation generated at the aberrant splice junction. In order
to quantitate A
42 and A
40 production, we used a previously
described specific ELISA (12). In this assay, two highly specific
monoclonal antibodies are used for the discrimination of both peptide
species (12). As reported before (12, 18, 26), the PS1
exon9 mutation results in an approximately 3-fold increase of the
A
42/A
total ratio (Fig. 2B). In contrast,
the cell line stably overexpressing PS1
exon9 C290S showed no
increased A
42/A
total ratio (Fig. 2B). Therefore using two different approaches we can show that the pathological activity of PS1
exon9 is independent of its lack of
proteolytic processing but is rather caused by the single amino acid
change at the aberrant splice junction.

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Fig. 2.
The pathological activity of
PS1 exon9 is due to a point mutation at codon
290. A, A production of K293 cells expressing
endogenous presenilins (control (Ctrl.)) and cell lines
overexpressing PS1 exon9 or PS1 exon9 C290S. Conditioned medium
from metabolically labeled kidney 293 cells expressing the indicated
presenilin variants was immunoprecipitated with antibody 6E10. A 40
and A 42 were separated on a previously described gel system, which
allows the specific resolution of both species (25). As reported
before, A 42 migrates faster as A 40 (25). Note that cells
expressing PS1 exon9 produced increased amounts of A 42, whereas
cells expressing PS1 exon9 C290S do not show such an increase. Two
individual immunoprecipitations of each cell lysate are shown.
B, quantitation of the A 42 and A 40 concentrations
in conditioned medium of kidney 293 cells expressing the indicated
presenilin variants using a previously described highly specific ELISA
(12). Expression of PS1 exon9 but not the expression of PS1 exon9
C290S results in a 3-fold increase of A 42 production. Identical
results were observed with independent cell clones.
|
|
Having demonstrated that PS1
exon9 C290S is pathologically inactive,
we also wanted to test if this protein is sufficient to rescue a
lin-12-mediated signaling defect that is a result of a
mutant PS homologue (sel-12) in Caenorhabditis
elegans (23). As reported previously, transgenic expression of
human presenilins rescues the phenotype caused by mutations of
sel-12 (22, 24). In contrast, transgenic expression of the
PS1
exon9 variant in C. elegans resulted in an incomplete
rescue of the sel-12 mutant phenotype, as indicated by a
significantly reduced brood size and reduced egg laying (Refs. 22 and
24; see also Table I). In order to assess
the in vivo function of PS1
exon9 C290S, we tested its
ability to rescue the putative sel-12 null allele
ar171 (23). Four independent transgenic strains expressing
PS1
exon9 C290S displayed robust egg laying (Table I). Based on both
numbers of laid progeny and numbers of eggs in utero, the
phenotype of these strains is almost indistinguishable of that of wild
type worms (Table I). These results demonstrate that PS1
exon9 C290S rescues the egg laying phenotype of the sel-12(ar171) mutant
animals significantly better than PS1
exon9 (Table I). Therefore, the reduced biological activity of PS1
exon9 is also due to a single point mutation and completely independent of the lack of proteolytic processing.
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Table I
Rescue of the sel-12 egg laying defect by human PS genes expressed from
the sel-12 promoter
For 50 transgenic animals each, the numbers of progeny were counted and
grouped in the following categories: +++, over 30 progeny laid by
individual animal; ++, 15-30 progeny laid; +, 5-15 progeny laid; ,
no progeny laid. The sel-12(ar171) strains carried an
additional unc-1(e538) marker that did not affect egg laying
or rescuing frequency. It was backcrossed several times from the strain
originally published (23).
|
|
 |
DISCUSSION |
Our results demonstrate that the pathological effect of the
PS1
exon9 mutation is independent of its lack of proteolytic cleavage and the deletion of the exon 9-encoded domain. Reverting the cysteine residue generated by the exon 8/10 splice junction at codon 290 back to
its wt residue (serine) still inhibits its proteolytic processing,
causes an accumulation of the uncleaved protein, and prevents formation
of endogenous PS fragments. Although the artificially generated
PS1
exon9 C290S variant behaves like PS1
exon9 in regard to the
characteristic biochemical features described above, it does not allow
pathological overproduction of A
42. Moreover, PS1
exon9 C290S
regains full biological activity in a very sensitive in vivo
assay system. This demonstrates that not only the pathological overproduction of A
42 but also the reduced biological function is
due to the single amino acid exchange. Therefore, similar to all other
known PS mutations, the pathological effect of the PS1
exon9 mutation
is due to a rather subtle amino acid exchange at a single highly
conserved codon whereas the large deletion of the complete domain
encoded by exon 9 does not affect the biological and pathological function. However, consistent with our previous results (27), we
suggest that PS molecules lacking the domain encoded by exon 9 mimic a
proteolytically processed and biologically active PS complex and
therefore rescue the phenotype of the mutant nematode. Consequently,
the rescuing activity of PS1
exon9 is significantly enhanced when the
pathologically relevant point mutation at codon 290 is reverted to the
wt residue.
It remains to be shown if the mutation at codon 290 is also
pathologically active within the full-length protein like all other
AD-associated point mutations. It may however be possible that the
mutation only exhibits a pathological activity if it is aberrantly
flanked by the domains encoded by exons 8 and 10. Structural changes
that may specifically occur in the PS1
exon9 molecule (12, 19) might
be responsible for the pathological activity of this very unusual mutation.
Based on our results it would be interesting to investigate if the
point mutations in the PS genes require the endoproteolytic cleavage
for their pathological activity. Therefore artificial PS molecules
should be generated containing an AD associated mutation as well as the
smallest possible alteration of the sequence at the cleavage site which
would inhibit PS processing. However, such mutations might be very
difficult to generate since PS can be cleaved at multiple sites (10,
12) and PS molecules containing larger deletions at the cleavage sites
might mimic a proteolytically processed PS molecule (27).
 |
ACKNOWLEDGEMENTS |
We thank Roland Donhauser for injecting PS
constructs into C. elegans and Drs. Bernard Lakowski and
Helmut Jacobsen for critical discussion.
 |
Note Added in Proof |
While this manuscript was in press, we
found that expression of PS1
exon9 C290S containing a FAD-associated
mutation (M146L) results in elevated A
42 production, in further
support of the findings presented here.
 |
FOOTNOTES |
*
This work was supported by grants from the Verum Foundation
(A
42 generation) and the Deutsche Forschungsgemeinschaft (DFG) (to
C. H. and R. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
**
To whom correspondence should be addressed: Central Institute of
Mental Health, Dept. of Molecular Biology, J5, 68159 Mannheim, Germany.
Tel.: 49-621-1703-884; Fax: 49-621-23429; E-mail: Haass{at}as200.zi-mannheim.de (for C. H.) or Genzentrum,
Feodor-Lynen-Str.25, 81377 Munich, Germany. Tel.: 49-89-7401-7347;
Fax: 49-89-7401-7314; E-mail: bmeister{at}LMB.uni-muenchen.de (for
R. B., for correspondence regarding C. elegans).
 |
ABBREVIATIONS |
The abbreviations used are:
AD, Alzheimer's
disease;
A
, amyloid
-peptide;
APP,
amyloid precursor
protein;
PS, presenilin;
wt, wild type;
PCR, polymerase chain reaction;
MEM, minimal essential medium;
ELISA, enzyme-linked immunosorbent
assay.
 |
REFERENCES |
-
Sherrington, R.,
Rogaev, E. I.,
Liang, Y.,
Rogaeva, E. A.,
Levesque, G.,
Ikeda, M.,
Chi, H.,
Lin, C.,
Li, G.,
Holman, K.,
Tsuda, T.,
Mar, L.,
Foncin, J.-F.,
Bruni, A. C.,
Montesi, M. P.,
Sorbi, S.,
Rainero, I.,
Pinessi, L.,
Nee, L.,
Chumakov, I.,
Pollen, D.,
Brookes, A.,
Sanseau, P.,
Polinsky, R. J.,
Wasco, W.,
da Silva, H. A. R.,
Haines, J. L.,
Pericak-Vance, M. A.,
Tanzi, R. E.,
Roses, A. D.,
Fraser, P. E.,
Rommens, J. M.,
and St. George-Hyslop, P. H.
(1995)
Nature
375,
754-760[CrossRef][Medline]
[Order article via Infotrieve]
-
Levy-Lahad, E.,
Wasco, W.,
Poorkaj, P.,
Romano, D. M.,
Oshima, J.,
Pettingell, W. H., Yu, C.,
Jondro, P. D.,
Schmidt, S. D.,
Wang, K.,
Crowley, A. C.,
Fu, Y.-H.,
Guenette, S. Y.,
Galas, D.,
Nemens, E.,
Wijsman, E. M.,
Bird, T. D.,
Schellenberg, G. D.,
and Tanzi, R. E.
(1995)
Science
269,
973-977[Medline]
[Order article via Infotrieve]
-
Rogaev, E. I.,
Sherrington, R.,
Rogaeva, E. A.,
Levesque, G.,
Ikeda, M.,
Liang, Y.,
Chi, H.,
Lin, C.,
Holman, K.,
Tsuda, T.,
Mar, L.,
Sorbi, S.,
Nacmias, B.,
Piacentini, S.,
Amaducci, L.,
Chumakov, I.,
Cohen, D.,
Lannfelt, L.,
Fraser, P. E.,
Rommens, J. M.,
and St. George-Hyslop, P. H.
(1995)
Nature
376,
775-778[CrossRef][Medline]
[Order article via Infotrieve]
-
Price, D.,
and Sisodia, S.
(1998)
Annu. Rev. Neurosci.
21,
479-505[CrossRef][Medline]
[Order article via Infotrieve]
-
Selkoe, D. J.
(1996)
J. Biol. Chem.
271,
18295-18298[Free Full Text]
-
Jarret, J. T.,
and Lansbury, P. T., Jr.
(1993)
Cell
73,
1055-1058[Medline]
[Order article via Infotrieve]
-
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]
-
Perez-Tur, J.,
Froehlich, S.,
Prihar, G.,
Crook, R.,
Baker, M.,
Duff, K.,
Wragg, M.,
Busfield, F.,
Lendon, C.,
Clark, R. F.,
Roques, P.,
Fuldner, R. A.,
Johnston, J.,
Cowburn, R.,
Forsell, C.,
Axelman, K.,
Lilius, L.,
Houlden, H.,
Karran, E.,
Roberts, G. W.,
Rossor, M.,
Adams, M. D.,
Hardy, J.,
Goate, A.,
Lannfelt, L.,
and Hutton, M.
(1995)
Neuroreport
7,
297-301[Medline]
[Order article via Infotrieve]
-
Thinakaran, G.,
Borchelt, D. R.,
Lee, M. K.,
Slunt, H. H.,
Spitzer, L.,
Kim, G.,
Ratovitsky, T.,
Davenport, F.,
Nordstedt, C.,
Seeger, M.,
Hardy, J.,
Levey, A. I.,
Gandy, S. E.,
Jenkins, N. A.,
Copeland, N. G.,
Price, D. L.,
and Sisodia, S. S.
(1996)
Neuron
17,
181-190[Medline]
[Order article via Infotrieve]
-
Podlisny, M.,
Citron, M.,
Amarante, P.,
Sherrington, R.,
Xia, W.,
Zhang, J.,
Diehl, T.,
Levesque, G.,
Fraser, P.,
Haass, C.,
Koo, E.,
Seubert, P.,
St. George-Hyslop, P.,
Teplow, D.,
and Selkoe, D.
(1997)
Neurobiol. Dis.
3,
325-337[CrossRef][Medline]
[Order article via Infotrieve]
-
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[Abstract/Free Full Text]
-
Steiner, H.,
Capell, A.,
Pesold, B.,
Citron, M.,
Kloetzel, P.-M.,
Selkoe, D.,
Romig, H.,
Mendla, K.,
and Haass, C.
(1998)
J. Biol. Chem.
273,
32322-32331[Abstract/Free Full Text]
-
Lee, M. L.,
Borchelt, D. R.,
Kim, G.,
Thinakaran, G.,
Slunt, H.,
Ratovitski, T.,
Martin, L. J.,
Kittur, A.,
Gandy, S.,
Levey, A.,
Jenkins, N.,
Copeland, N.,
Price, D. L.,
and Sisodia, S.
(1997)
Nat. Med.
3,
756-760[Medline]
[Order article via Infotrieve]
-
Mercken, M.,
Takahashi, H.,
Honda, T.,
Sato, K.,
Murayama, M.,
Nakazato, Y.,
Noguchi, K.,
Imahori, K.,
and Takashima, A.
(1996)
FEBS Lett.
389,
297-303[CrossRef][Medline]
[Order article via Infotrieve]
-
Murayama, O.,
Honda, T.,
and Mercken, M.
(1997)
Neurosci. Lett.
229,
61-64[CrossRef][Medline]
[Order article via Infotrieve]
-
Grünberg, J.,
Capell, A.,
Leimer, U.,
Steiner, B.,
Steiner, H.,
Walter, J.,
and Haass, C.
(1997)
Alzheimer's Res.
3,
253-259
-
Citron, M.,
Oltersdorf, T.,
Haass, C.,
McConlogue, A. Y.,
Seubert, P.,
Vigo-Pelfrey, C.,
Lieberburg, I.,
and Selkoe, D. J.
(1992)
Nature
360,
672-674[CrossRef][Medline]
[Order article via Infotrieve]
-
Citron, M.,
Westaway, D.,
Xia, W.,
Carlson, G.,
Diehl, T. S.,
Levesque, G.,
Johnson-Wood, K.,
Lee, M.,
Seubert, P.,
Davis, A.,
Kholodenko, D.,
Motter, R.,
Sherrington, R.,
Perry, B.,
Yao, H.,
Strome, R.,
Lieberburg, I.,
Rommens, J.,
Kim, S.,
Schenk, D.,
Fraser, P.,
St. George-Hyslop, P.,
and Selkoe, D.
(1997)
Nat. Med.
3,
67-72[Medline]
[Order article via Infotrieve]
-
Walter, J.,
Grünberg, J.,
Capell, A.,
Pesold, B.,
Schindzielorz, A.,
Citron, M.,
Mendla, K.,
St. George-Hyslop, P.,
Multhaup, G.,
Selkoe, D. J.,
and Haass, C.
(1997)
Proc. Natl. Acad. Sci. U. S. A.
94,
5349-5354[Abstract/Free Full Text]
-
Walter, J.,
Grünberg, J.,
Schindzielorz, A.,
and Haass, C.
(1998)
Biochemistry
37,
5961-5967[CrossRef][Medline]
[Order article via Infotrieve]
-
Haass, C.,
Schlossmacher, M.,
Hung, A. Y.,
Vigo-Pelfrey, C.,
Mellon, A.,
Ostaszewski, B.,
Lieberburg, I.,
Koo, E.,
Schenk, D.,
Teplow, D.,
and Selkoe, D. J.
(1992)
Nature
359,
322-325[CrossRef][Medline]
[Order article via Infotrieve]
-
Baumeister, R.,
Leimer, U.,
Zweckbronner, I.,
Jakubek, C.,
Grünberg, J.,
and Haass, C.
(1997)
Genes Funct.
1,
149-159[Medline]
[Order article via Infotrieve]
-
Levitan, D.,
and Greenwald, I.
(1995)
Nature
377,
351-354[CrossRef][Medline]
[Order article via Infotrieve]
-
Levitan, D.,
Doyle, T.,
Brousseau, D.,
Lee, M.,
Thinakaran, G.,
Slunt, H.,
Sisodia, S.,
and Greenwald, I.
(1996)
Proc. Natl. Acad. Sci. U. S. A.
93,
14940-14944[Abstract/Free Full Text]
-
Klafki, H.-W.,
Wilfang, J.,
and Staufenbiel, M.
(1996)
Anal. Biochem.
237,
24-29[CrossRef][Medline]
[Order article via Infotrieve]
-
Borchelt, D.,
Thinakaran, G.,
Eckman, C.,
Lee, M.,
Davenport, F.,
Ratovitsky, T.,
Prada, C.-M.,
Kim, G.,
Seekins, S.,
Yager, D.,
Slunt, H.,
Wang, R.,
Seeger, M.,
Levey, A.,
Gandy, S.,
Copeland, N.,
Jenkins, N.,
Price, D.,
Younkin, S.,
and Sisodia, S. S.
(1996)
Neuron
17,
1005-10013[Medline]
[Order article via Infotrieve]
-
Capell, A.,
Grünberg, J.,
Pesold, B.,
Diehlmann, A.,
Citron, M.,
Nixon, R.,
Beyreuther, K.,
Selkoe, D.,
and Haass, C.
(1998)
J. Biol. Chem.
273,
3205-3211[Abstract/Free Full Text]
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.