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J Biol Chem, Vol. 274, Issue 46, 32997-33001, November 12, 1999
,From the Institut für Zellbiochemie und Klinische Neurobiologie, Universitätsklinikum Hamburg-Eppendorf, Universität Hamburg, Martinistrasse 52, 20246 Hamburg, Germany
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ABSTRACT |
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 TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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By using the yeast two-hybrid system we
identified a novel protein from the human brain interacting with the C
terminus of somatostatin receptor subtype 2. This protein termed
somatostatin receptor interacting protein is characterized by a novel
domain structure, consisting of six N-terminal ankyrin repeats followed by SH3 and PDZ domains, several proline-rich regions, and a C-terminal sterile Targeting of neurotransmitter receptors to postsynaptic or
presynaptic sites is an area that has been widely studied in recent years; a large body of evidence has accumulated showing that many receptors are anchored at their specifc site of action by specialized anchoring proteins, which may link receptors to components of the
synaptic structure or the cytoskeleton (1, 2). This is true for
inhibitory as well as excitatory receptors of the family of
ligand-gated ion channels. For the second large family of
neurotransmitter receptors, the seven transmembrane domain G-protein-coupled receptors, only very recently have some proteins been
identified that may be involved in anchoring or linkage to the
cytoskeleton. These include the homer proteins, which are tightly
associated with metabotropic glutamate receptors via a PDZ1 domain in homer and the
C terminus of the mGluRs (3). However, for the large majority of
G-protein-coupled receptors, no intracellular associated proteins have
been identified so far beyond those proteins which are necessary for
signal transduction and functional regulation of the receptors,
i.e. the G-proteins and proteins of the arrestin family
(4).
We have begun to address this issue for members of the somatostatin
receptor family (SSTRs). SSTRs are widely expressed in neuronal tissue
and modulate synaptic responses by interacting with inhibitory
G-proteins in presynaptic as well as postsynaptic compartments of
neurons (e.g. Refs. 5-8). Recently we have used the yeast
two-hybrid system to screen for proteins intracellularly associated
with SSTR2, one of the major SSTR subtypes in the mammalian brain. Here
we show that the C terminus of SSTR2, which contains the consensus
sequence for recognition of PDZ domains, interacts with a novel protein
termed SSTR interacting protein or SSTRIP. The latter defines a novel
family of multidomain cytoskeletal anchoring proteins that are highly
enriched in the postsynaptic density fraction derived from rat brain.
cDNA Cloning--
A yeast two-hybrid screen using the rat
SSTR2 C terminus as a bait in the Gal4-DNA binding domain vector pAS2
was described previously (9). The initial partial clone of 900 base
pairs (clone 16; Ref. 9) coding for the PDZ domain of SSTRIP was used
as a probe to screen human fetal brain, adult thalamus, and hippocampus
cDNA libraries (CLONTECH) in order to obtain
the full-length sequence of SSTRIP. By using oligonucleotide primers
based on the human sequence, a partial sequence was also amplified from rat hypothalamic cDNA and used for screening of a rat brain
cDNA library (kindly provided by Dr. Rainer Reinscheid).
Overlay Assay--
A cDNA fragment encoding the PDZ domain
of human SSTRIP was cloned into the GST fusion protein vector pGEX2T
(Amersham Pharmacia Biotech) and used for the generation of fusion
protein. Overlay assays with a biotinylated fusion protein containing
the C terminus of SSTR2 were performed as described (9).
Expression in HEK Cells, Immunoprecipitation--
The
construction of an SSTR2-cDNA carrying an N-terminal T7 epitope tag
in the expression vector pcDNA3 (Invitrogen, Leek, The Netherlands)
has been described before (10, 11). Ntag-SSTR2 and SSTRIP were
coexpressed in human embryonic kidney (HEK) cells by transient
transfection using the calcium phosphate method as described (12). For
immunoprecipitations, cells were lyzed in 1 ml of RIPA buffer (1%
Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, 150 mM
NaCl, 50 mM Tris-HCl, pH 7.4) on ice for 20 min and
centrifuged at 15,000 × g for 20 min. The
epitope-tagged receptor was precipitated from the supernatant fraction
using the monoclonal T7-antibody (3 µg) and protein A-Sepharose
(Amersham Pharmacia Biotech; 100 µl of a 50% suspension in RIPA
buffer) as described (11). Precipitates were washed five times with
RIPA buffer and denatured by boiling in Laemmli sample buffer. After
separation by polyacrylamide gel electrophoresis, proteins were blotted
onto nitrocellulose membranes. SSTRIP was then detected by
immunoblotting using the anti-PDZ domain antibody as a primary antibody
and goat anti-rabbit coupled to alkaline phosphatase as secondary antibody.
In Situ Hybridization--
Whole brains of adult rats were
frozen on dry ice, and 20-µm sections were prepared on a cryostat
(Leitz, Wetzlar, Germany). Antisense RNA probes labeled with
Fractionation of Rat Brain Membranes--
Rat brain membranes
were fractionated according to the method described by Carlin et
al. (15). Briefly, a postnuclear membrane fraction (crude
membranes) was applied to a sucrose multistep gradient, and the
synaptosomal fraction was recovered at the interface between 1.0 and
1.2 M sucrose after centrifugation at 82,500 × g for 2 h. After extraction with 1% Triton X-100, the
PSD-one Triton fraction was obtained in a second ultracentrifugation
step at 200,000 × g at the interface between 1.5 and
2.0 M sucrose and used for Western blotting experiments.
Specific Interaction between a Novel PDZ Domain Protein and the C
Terminus of the Rat SSTR2--
We used the C terminus of the rat SSTR2
as a bait in a yeast two-hybrid screen for potentially interacting
proteins (9); one novel clone of 900 base pairs, termed SSTRIP for
somatostatin receptor interacting protein, was obtained from a human
cDNA library which interacted specifically with SSTR2 when compared
with the empty bait vector or nonrelated control proteins. This clone
encodes a protein fragment that contains a PDZ domain which is highly similar to that of cortactin-binding protein 1 (CortBP1; see Refs. 9
and 16).
Screening of rat and human brain cDNA libraries yielded several
splice variants of the full-length SSTRIP as depicted in Fig. 1. The longest cDNA derived from
human brain contains an open reading frame of 6546 base pairs, coding
for a protein of 240 kDa. In this protein, the PDZ domain is preceded
by the SH3 domain and a domain containing six ankyrin repeats. The PDZ
domain is followed by seven proline-rich domains that may function as
SH3-binding domains (17). At the C terminus a sterile
A splice variant of SSTRIP was observed in which the N-terminal region
with the six ankyrin repeats as well as the SH3 domain is replaced by
the alternative N terminus labeled as exon a in Fig. 1.
Accordingly, a major splice site appears to be located between the SH3
domain and the PDZ domain, as an additional partial clone was observed
which contains the SH3 and the PDZ domains but lacks an insert of 9 amino acids (insert b in Fig. 1) at this position.
Alternative splicing C-terminal of the PDZ domain leads to transcripts
that contain an additional insert of 8 amino acids (exon c
in Fig. 1) between the PDZ and the first proline-rich domain. However,
this variant was so far detected only in rat tissues.
In order to confirm the interaction observed in the yeast system in
terms of true protein-protein interactions, we performed an overlay
assay with a GST fusion protein of the PDZ domain of SSTRIP. Fusion
proteins were separated by SDS-polyacrylamide gel electrophoresis and
blotted onto nitrocellulose membranes; a biotinylated fusion protein of
the SSTR2 C terminus was used as a probe, according to Ref. 9. In these
experiments, the PDZ domain of SSTRIP exhibited a strong interaction
with SSTR2, whereas the GST control protein did not react (Fig.
2A). In vivo
attempts to coimmunoprecipitate SSTR2 and SSTRIP from a plasma membrane
fraction of rat brains were not successful due to the fact that SSTRIP
could only be solubilized in the presence of RIPA supplemented with 2%
SDS. These conditions did not permit the detection of specific
interactions between SSTR2 and SSTRIP although immunoreactivity of the
individual proteins was still present in the solubilized membrane
fraction (data not shown). To circumvent this technical problem
in vitro, coexpression experiments were performed in HEK
cells. An expression vector containing the N-terminal part of SSTRIP
(amino acids 1-1288), including the ankyrin, SH3, PDZ, and some of the
proline-rich regions, was cotransfected with the rat SSTR2 carrying a
N-terminal T7 epitope tag (NT7-SSTR2). When SSTR2 was precipitated from
cellular lysates using the T7 antibody, SSTRIP could be efficiently
detected in the precipitate by Western blotting. SSTRIP could not be
detected in the precipitate when SSTR2 was omitted from the
transfection mixture, indicating that SSTR2 specifically interacts with
SSTRIP in these cells (Fig. 2B). It should be noted that
SSTRIP coexpressed with the inward rectifier potassium channel protein
Kir 2.1 known to carry the consensus sequence for the PDZ domain and to
bind to PSD95 (20) in HEK cells did not show coimmunoprecipitation from
cell lysates (data not shown) excluding the possibility that SSTRIP
interacts with any protein carrying the PDZ consensus sequence at its C
terminus.
The specificity of the interaction between the SSTR2 C terminus and the
PDZ domain of SSTRIP was further analyzed in the yeast two-hybrid
system. Bait plasmids encoding the C termini of SSTR1-4 as well as a
C-terminally mutated SSTR2 (the last amino acid residue isoleucine is
replaced by seven amino acids derived from the bait vector sequence)
were coexpressed with the initial target vector obtained from the human
brain cDNA library. Activation of the His-3 reporter gene that
triggers growth on His-deficient media was observed only in the
presence of the wild-type SSTR2 and, to a lesser degree, in the
presence of the SSTR4 C terminus (Table I). No growth was seen when the SSTR2 C
terminus was modified. These data were confirmed by a SSTRIP Is Expressed in Multiple Human Brain Tissues--
As
assessed by Northern blot analysis of RNA from various human organs,
the SSTRIP gene is expressed in the human brain, particularly in the
amygdala, hippocampus, substantia nigra, and thalamus; there two
transcripts of 9 and 7.5 kb are detected (Fig.
3), the latter probably corresponding to
one of the splice variants of SSTRIP shown in Fig. 1. Expression in
organs other than brain such as heart, skeletal muscle, kidney, or
liver is restricted to the shorter transcript; this may indicate that
shorter splice variants of SSTRIP are ubiquitously expressed in tissues
other than brain.
In order to compare the expression pattern of SSTR2 and SSTRIP, we
performed in situ hybridization experiments of rat brain sections. As shown in Fig. 4, in general
a striking overlap in the expression pattern of both SSTR2 and SSTRIP
was observed, including all layers of the cortex and the hippocampus.
Both proteins are also prominently expressed in the medial habenula. A
major difference between SSTR2 and SSTRIP was seen in the hippocampus; here both mRNAs are seen in the CA1 and CA2 regions; however, the
SSTRIP mRNA was detected not only in cell bodies but also in the
molecular layer, suggesting that SSTRIP might add to the growing number
of mRNAs which are transported into neuronal dendrites (Fig. 4).
SSTRIP Is Enriched in the Postsynaptic Density--
A fusion
protein containing the PDZ domain was used to raise rabbit polyclonal
antibodies; due to the sequence similarity between CortBP1 and the
SSTRIP protein in this domain, these antibodies recognize both proteins
from rat and human tissues with similar efficiency. This antiserum was
used in Western blotting experiments on rat brain membrane fractions.
Several specific bands were observed at molecular masses ranging from
180 to 260 kDa (Fig. 5). CortBP1 from rat
brain migrates at an apparent molecular mass of 180 kDa (16),
suggesting that the multiple bands at higher molecular weights
correspond to the various splice variants of SSTRIP. Specific labeling
for CortBP as well as for SSTRIP was obtained in a crude membrane
fraction as well as in the synaptosomal fraction and in postsynaptic
densities (so-called PSD-one Triton fraction). All bands are strongly
enriched in the PSD-one Triton fraction when compared with the crude
membrane fraction and the synaptosomal fraction. This enrichment is
similar to that observed in a parallel experiment with an antibody
specific for synapse-associated protein 102 (SAP102), which has been
characterized as a major constituent of postsynaptic densities (24). To
verify the observation that SSTRIP is enriched in the post-synaptic
density fraction, we used a second antibody directed against the
ankyrin repeat domain of SSTRIP. In this experiment only a single
significant band at 240 kDa was observed which is also strongly
enriched in the PSD fraction (Fig. 5). Thus our data show that the
ankyrin/SH3 domain of SSTRIP is contained in one of the several
variants of the SSTRIP/CortBP1 family, all of which are enriched in the
postsynaptic density fraction.
Here we report the cloning of human SSTRIP, a protein that
interacts specifically with the C terminus of the SSTR2 via a PDZ domain-type interaction. During the cloning work of SSTRIP, two sequences from rat brain termed synamon, Shank1 and Spank1, have been
deposited in the GenBankTM data base which exhibit strong
homology to the human SSTRIP protein and are in fact identical to our
partial clone from rat. Rat Shank1 and Spank1 exhibit a domain
structure identical to human SSTRIP, whereas the reported rat synamon
sequence lacks the SAM domain. In addition, a protein termed ProSAP1
(proline-rich synapse associated protein 1) was reported (25) which is
largely identical to CortBP1 but which exhibits several alternative
splicing sites in positions homologous to those reported here for
SSTRIP. The homology between SSTRIP and CortBP1/ProSAP1 is limited to
the PDZ and SAM domains, where the sequence identity between the two
proteins reaches 86 (PDZ) and 73% (SAM). In contrast, the region
between the PDZ and the SAM domain displays a sequence identity of 25%
only. A third cDNA may be present, as both SSTRIP and
CortBP1/ProSAP1 align to a genomic fragment from the human chromosome
22, with very high sequence similarity again in the PDZ and SAM
domains. As the PDZ domains of these three proteins are structurally
very similar, it appears likely that the many variants of these three proteins may be responsible for anchoring SSTR2 in many different localizations in neurons. This is in agreement with immunohistochemical data, which have shown that the SSTR2 protein is present in presynaptic as well as in somatodendritic, postsynaptic compartments of neuronal cells (6, 8).
The common domain structure and splicing pattern seen when comparing
SSTRIP and CortBP1/ProSAP1 (9, 16, 25) suggests that these three
proteins constitute a new family of multidomain proteins. It remains to
be shown, however, if the ankyrin repeat domain and the SH3 domain are
also present in splice variants of CortB1 and the unknown protein
derived from human chromosome 22. The strong enrichment of proteins of
this family in the postsynaptic density fraction from rat brain
indicates that we have detected an important structural component of
this postsynaptic structure. In this respect the function of SSTRIP and
its homologs may go far beyond an anchoring function for somatostatin
receptors, as family members have been identified independently by
experimental approaches looking for interaction partners for cortactin
(16), or the SAP-associated protein/guanylate kinase-associated protein (see annotations for the rat homologs of SSTRIP in the
GenBankTM data base; see Refs. 26 and 27). Similarly,
multiple interaction partners have been found for the PDZ domains of
the PSD-95/SAP102 family of proteins (28), which bind glutamate
receptor as well as potassium channel subunits and probably many other
proteins. Further yeast two-hybrid experiments will help to identify
additional proteins that complex with the PDZ domains as well as with
the ankyrin, SH3, and SAM domains of SSTRIP.
motif. It consists of 2185 amino acid residues encoded by a
9-kilobase pair mRNA; several splice variants have been detected in
human and rat cDNA libraries. Sequence comparison suggests that the
novel multidomain protein, together with cortactin-binding protein,
forms a family of cytoskeletal anchoring proteins. Fractionation of rat
brain membranes indicated that somatostatin receptor interacting protein is enriched in the postsynaptic density fraction. The interaction of somatostatin receptor subtype 2 with its interacting protein was verified by overlay assays and coimmunoprecipitation experiments from transfected human embryonic kidney cells. Somatostatin receptor subtype 2 and the interacting protein display a striking overlap of their expression patterns in the rat brain. Interestingly, in the hippocampus the mRNA for somatostatin receptor interacting protein was not confined to the cell bodies but was also observed in
the molecular layer, suggesting a dendritic localization of this mRNA.
INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
MATERIALS AND METHODS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
-35S-UTP (NEN Life Science Products) were generated
using the rat SSTR2 cDNA (13) and a rat SSTRIP-clone matching bases
1544-2619 of the human SSTRIP sequence. In situ
hybridization experiments were performed as described (14). Sections
were exposed to BioMax MR film for 3 days. For higher resolution
studies, the same slides were dipped into Kodak NTB-2 nuclear emulsion,
developed in Dektol developer (Eastman Kodak Co.) after 2-3 weeks of
exposure, and subsequently stained with Giemsa (Sigma).
RESULTS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
motif (SAM
domain) is observed which has been implicated as a dimerization motif for transcription factors as well as receptor tyrosine kinases (18,
19).
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Fig. 1.
Structure of human SSTRIP. A,
amino acid sequence. The sequence of the longest form of SSTRIP is
shown. In light gray with black letters, the
ankyrin repeat region (Ank), in black with
white letters the SH3 domain (SH3), the PDZ
domain (PDZ), and the SAM domain (SAM), in dark
gray with white letters, proline-rich regions (PR1-7)
are depicted. The alternatively spliced insert b is
boxed; the sequence of the alternative N terminus
(a) which may be introduced at this position is shown at the
bottom of A. A further splice site is indicated
by an arrowhead where the segment c may be
introduced. This segment was, however, only detected in cDNA clones
obtained from rat tissues; the flanking sequences of this splice site
are almost identical in rat and human SSTRIP. The sequences of
alternative exons a, b, and c are shown at the
bottom in bold letters, with the flanking
sequences in the mature protein in normal letters. B, domain
structure. Ank6 denotes a set of six ankyrin repeats; the
shaded bars indicate the position of proline-rich, putative
SH3 binding domains. All domains were identified by sequence analysis
using the ISREC Profile Scan server maintained by the Swiss Institute
for experimental cancer research. The positions where the alternative
exons a, b, and c are introduced are indicated.
Map not drawn to scale.
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Fig. 2.
Interaction of SSTRIP and SSTR2.
A, overlay assay. A GST fusion protein of the PDZ domain of
SSTRIP and a GST control protein were run on SDS-polyacrylamide gel
electrophoresis and either stained with Coomassie Brilliant Blue
(right panel) or blotted onto nitrocellulose membranes and
probed with the biotinylated fusion protein containing the SSTR2 C
terminus (left panel). Specifically bound fusion protein was
detected using alkaline phosphatase coupled to avidin. GST,
lane containing only the GST carrier protein; SSTRIP, amino
acid residues 655-796 of full-length human SSTRIP. B,
immunoprecipitation from transfected HEK cells. HEK cells were
transiently transfected with an expression vector for a truncated
(trunc.) version of human SSTRIP (amino acid residue
1-1288) either alone or in combination with Ntag-SSTR2. The 1st
two lanes from the left show Western blots from
cellular lysates using the 189.3 antiserum that recognizes SSTRIP. The
two lanes from the right show Western blots of
immunoprecipitates where the SSTR2 was precipitated from cellular
lysates using the T7 antibody directed against the epitope tag at
the N terminus of the receptor. Note the absence of staining in lanes
where SSTR2 cDNA was omitted from the transfection mixture.
IP, immunoprecipitate.
-galactosidase
assay on His-positive colonies, indicating that the interaction between
SSTR2 and the PDZ domain requires an intact C terminus (Table I). A
possible interaction of SSTR4 with the SSTRIP-PDZ domain that was
weakly detected here could not be confirmed in an overlay experiment (data not shown).
SSTRIP specifically interacts with the SSTR2 C terminus
, no
growth; +++, the highest number of colonies were observed with the
SSTR2 construct; +, very few colonies were obtained with the SSTR4
construct. ND, not determined. The respective references are given in
parentheses.
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Fig. 3.
Expression of the SSTRIP gene in human
tissues. Human brain multiple tissue (upper panels) and
human multiple tissue (lower panels; CLONTECH) Northern
blots were hybridized with 32P-labeled probes specific for
human SSTRIP and actin. The exposure times were as follows: SSTRIP,
top, 4 h; bottom, 3 days; actin: 1 h. Two
significant bands (7 and 9 kb) were observed in various brain regions,
whereas only the lower band was observed in other tissues. Lanes
are as follows: 1, amygdala; 2, caudate
nucleus; 3, corpus callosum; 4, hippocampus;
5, total brain; 6, substantia nigra;
7, subthalamic nucleus; 8, thalamus;
9, brain; 10, heart; 11, skeletal
muscle; 12, colon; 13, thymus; 14,
spleen; 15, kidney; 16, liver; 17,
small intestine; 18, placenta; 19, lung;
20, peripheral blood leukocytes.
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Fig. 4.
Comparison of the expression patterns of
SSTR2 and SSTRIP in the rat brain. Adjacent brain sections were
probed with 35S-labeled antisense riboprobes for the rat
SSTR2 (A and C) and rat SSTRIP (B, D,
F, and H). Shown are coronal (A, B, E, F, G,
and H) and sagital sections (C and D)
prepared from frozen rat brains. E-H show high resolution
analysis of medial habenula (E and F) and
hippocampus (G and H) probed with radiolabeled
SSTRIP. E and G are bright field images, and
F and H show the corresponding dark field
photomicrographs. 3V, third ventricle; CA, ammons
horn; Cb, cerebellum; Cx, cortex; DG,
dentate gyrus; Hi, hippocampus; MHb, medial
habenula; Pir, piriform cortex.
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Fig. 5.
SSTRIP is concentrated in rat brain membrane
fractions enriched for postsynaptic densities. Crude membranes
(c), synaptosomal (s), and PSD-one Triton
(p) fractions were prepared as described under "Materials
and Methods." 10 µg of each fraction were applied to
SDS-polyacrylamide gel electrophoresis and analyzed by Western blotting
using the 189.3 antiserum (directed against the PDZ domain of human
SSTRIP) preincubated with an excess of antigen, the 189.3 antiserum
alone, and an antiserum raised against the ankyrin domain (amino acids
212-399; A.3). A rabbit polyclonal antibody against synapse-associated
protein SAP102 (Müller et al. (24), right)
was used as a control for the enrichment of postsynaptic densities.
Open arrowheads point at bands at 260 and 180 kDa, which
probably represent the longest form of SSTRIP and CortBP1,
respectively; the closed arrowhead points at a cluster of
bands that probably represent additional PDZ domain containing splice
variants of SSTRIP and possibly the unknown protein derived from the
human chromosome 22 (see "Discussion"). All these bands are
specifically labeled as they are absent when the experiment is
performed in the presence of excess antigen.
DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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ACKNOWLEDGEMENTS |
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We thank Drs. Chica Schaller and Irm Hermans-Borgmeyer (Zentrum für Molekulare Neurobiologie, Hamburg, Germany) for help with the in situ hybridization experiments and Dr. Stefan Kindler for the SAP102-specific antibody.
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Note Added in Proof |
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During the revision of this manuscript, the rat homolog of SSTRIP has been published as Shank1 by Lim, S., Naisbitt, S., Yoon, J., Hwang, J.-I., Suh, P.-G., Sheng, M., and Kim, E. (1999) J. Biol. Chem. 274, 29510-29518 and Naisbitt, S., Kim. E., Tu, J. C., Xiao, B., Sala, S., Valtschanoff, J., Weinberg, R. J., Worley, P. F., and Sheng, M. (1999) Neuron 23, 569-582 and as Synamon by Yao, I., Yutaka, H., Hirao, K., Deguchi, M., Ide, N., Takeuchi, M., and Takai, Y. (1999) J. Biol. Chem. 274, 27463-27466.
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FOOTNOTES |
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* This work was supported by Deutsche Forschungsgemeinschaft Grant SFB545/B7 (to H.-J. K. and D. R.).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.
Presented this work as part of a thesis.
§ To whom correspondence should be addressed: Institut für Zellbiochemie und Klinische Neurobiologie, UKE, Martinistrasse 52, 20246 Hamburg, Germany. Tel.: 49 40 42803 3344; Fax: 49 40 42803 4541;E-mail: richter@uke.uni-hamburg.de.
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ABBREVIATIONS |
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The abbreviations used are:
PDZ, PSD-95/discs
large/ZO-1;
CortBP1, cortactin-binding protein 1;
GST, glutathione
S-transferase;
HEK, human embryonic kidney;
PSD, postsynaptic density;
SAM, sterile motif;
SAP102, synapse-associated protein 102;
SH3, Src homology 3;
SSTR, somatostatin
receptor;
SSTRIP, somatostatin receptor interacting protein;
kb, kilobase pairs.
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REFERENCES |
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TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
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