Abstract
PSD-95 is a component of postsynaptic densities in central synapses. It contains three PDZ domains that localize N -methyl- d -aspartate receptor subunit 2 (NMDA2 receptor) and K + channels to synapses. In mouse forebrain, PSD-95 bound to the cytoplasmic COOH-termini of neuroligins, which are neuronal cell adhesion molecules that interact with β-neurexins and form intercellular junctions. Neuroligins bind to the third PDZ domain of PSD-95, whereas NMDA2 receptors and K + channels interact with the first and second PDZ domains. Thus different PDZ domains of PSD-95 are specialized for distinct functions. PSD-95 may recruit ion channels and neurotransmitter receptors to intercellular junctions formed between neurons by neuroligins and β-neurexins.
References
39
Referenced
619
10.1016/0896-6273(92)90245-9
-
Kistner U., et al., J. Biol. Chem. 268, 4580 (1993);
(
10.1016/S0021-9258(18)53433-5
) / J. Biol. Chem. by Kistner U. (1993) -
Hunt C. A., Schenker L. J., Kennedy M. B., J. Neurosci. 16, 1380 (1996);
(
10.1523/JNEUROSCI.16-04-01380.1996
) / J. Neurosci. by Hunt C. A. (1996) -
Kim E., Cho K., Rothschild A., Sheng M., Neuron 17, 103 (1996);
(
10.1016/S0896-6273(00)80284-6
) / Neuron by Kim E. (1996) - ; B. M. Müller et al. ibid. p. 255.
10.1016/0092-8674(81)90009-X
-
Ruff P., Speicher D. W., Husain-Chishti A., Proc. Natl. Acad. Sci. U.S.A. 88, 6595 (1991);
(
10.1073/pnas.88.15.6595
) / Proc. Natl. Acad. Sci. U.S.A. by Ruff P. (1991) - Willott E., et al., ibid. 90, 7834 (1993); / ibid. by Willott E. (1993)
-
Woods D. F., Bryant P. J., J. Cell Sci. 17, 171 (1993);
(
10.1242/jcs.1993.Supplement_17.25
) / J. Cell Sci. by Woods D. F. (1993) -
Jesaitis L. A., Goodenough D. A., J. Cell Biol. 124, 949 (1994).
(
10.1083/jcb.124.6.949
) / J. Cell Biol. by Jesaitis L. A. (1994) 10.1126/science.7569905
-
Kim E., Niethammer M., Rothschild A., Jan Y. N., Sheng M., Nature 378, 85 (1995);
(
10.1038/378085a0
) / Nature by Kim E. (1995) -
Niethammer M., Kim E., Sheng M., J. Neurosci. 16, 2157 (1996);
(
10.1523/JNEUROSCI.16-07-02157.1996
) / J. Neurosci. by Niethammer M. (1996) -
Lau L.-F., et al., J. Biol. Chem. 271, 21622 (1996).
(
10.1074/jbc.271.35.21622
) / J. Biol. Chem. by Lau L.-F. (1996) 10.1016/0092-8674(95)90396-8
-
Ichtchenko K., Nguyen T., Südhof T. C., J. Biol. Chem. 271, 2676 (1996).
(
10.1074/jbc.271.5.2676
) / J. Biol. Chem. by Ichtchenko K. (1996) 10.1126/science.1621094
-
Ushkaryov Y. A., Südhof T. C., Proc. Natl. Acad. Sci. U.S.A. 90, 6410 (1993) ;
(
10.1073/pnas.90.14.6410
) / Proc. Natl. Acad. Sci. U.S.A. by Ushkaryov Y. A. (1993) -
Ushkaryov Y. A., et al., J. Biol. Chem. 259, 11987 (1994);
(
10.1016/S0021-9258(17)32671-6
) / J. Biol. Chem. by Ushkaryov Y. A. (1994) -
Ullrich B., Ushkaryov Y. A., Südhof T. C., Neuron 14, 497 (1995).
(
10.1016/0896-6273(95)90306-2
) / Neuron by Ullrich B. (1995) - T. Nguyen and T. C. Südhof J. Biol. Chem. in press.
- Total RNA from forebrain was isolated from mice on postnatal days 0 2 5 7 10 12 15 20 30 and 60. RNA blots obtained with formaldehyde gels were hybridized with 32 P-labeled probes from neurexins neuroligins GAP-43 syntaxin 1 synaptobrevin 2 and synaptotagmin 1. All blots were hybridized with a GAPDH probe to control for RNA loading. 32 P signals were quantified by PhosphorImager detection and are expressed as ratio to GAPDH.
- A rat brain cDNA library in the yeast-two hybrid prey vector pVP16 [
10.1016/0092-8674(93)90307-C
- ] was screened as described [
-
Fields S., Song O., Nature 340, 245 (1994);
(
10.1038/340245a0
) / Nature by Fields S. (1994) - ] (14) with the bait vector pBTM116-NL2-1 (8). Of 16 prey clones isolated pPrey500 encodes residues 1 to 287 and pPrey514 encodes residues 308 to 425 of PSD-95 (1). β-Galactosidase assays were corrected for protein concentration [M. D. Rose F. Winston P. Hieter Methods in Yeast Genetics (Cold Spring Harbor Laboratory Press Cold Spring Harbor NY 1990);
-
Hata Y., Südhof T. C., J. Biol. Chem. 270, 13022 (1995)].
(
10.1074/jbc.270.22.13022
) / J. Biol. Chem. by Hata Y. (1995) - The full-length sequence of rat PSD-95 (residues 1 to 724) its NH 2 -terminus (residues 1 to 431) and its COOH-terminus (residues 430 to 724) were cloned by PCR with oligonucleotides 1176 (GCGCTCGAGGTACCATGGACTGTCTCTGTATAGTGAC) 1177 (CGCGTCGACTAGAAGCCCCTCTTGGGGTT) 1178 (CGCCTCGAGGAATCAGAGTCTCTCTCGGGC) and 1198 (GCGAATTCTCCATGGGCTTCTACATTAGGGCC). To identify additional members of the PSD-95 family we used a degenerate PCR strategy. Products from PCRs with redundant oligonucleotide primers complementary to conserved sequences (T73 = TICCI- [C T]A[C T]ACIACI[C A]GICC; T75 = AC[G A] TCIA[G A]IAT[G A]CA[G A]TG; T76 = GGIGA[C T]ATI[C T]TICA[C T]GT; I = inosine; brackets = redundant positions) were subcloned and sequenced. We used two products (A290 and A309) containing unusual sequences related to PSD-95 as probes to isolate full-length clones pDLG54 and pDLG70 (GenBank accession numbers and ). The same two isoforms have been independently reported as PSD-93/chapsyn and SAP102 (1). pDLG54 is identical with SAP102 except for the absence of the sequence VTSNTSDSESSSKG (residues 627 to 630) (17) which suggests that pDLG54 represents a splice variant of SAP102. pDLG70 corresponds to PSD-93/chapsyn with several amino acid differences that may be due to alternative splicing or sequencing variations. The neuroligin bait vectors used encode the following residues of the rat proteins (4): pBTM116-NL1-1 718–843; pBTM116-NL1-2 718–840; pBTM116-NL1-5 718–828; pBTM116-NL1-6 828–843; pBTM116-NL1-10 796–843; pBTM116-NL2-1 699–836; pBTM116-NL2-2 699–786; pBTM116-NL2-3 786–836; pBTM116-NL3 730–848. The NMDA2 receptor bait plasmids encode residues 1455–1464 (pBTM116NMDAR2A) and 1473–1482 (pBTM116NMDAR2B) of the rat NMDA-2A and -2B receptors. pBTM116Shaker encodes residues 645–654 of the rabbit Kv1.4 K + channel isoform. The different prey vectors are from rat except for ZO-1 (mouse) and dlg-A ( Drosophila ) and encode the following residues: PSD-95 pVP16PSD-95-2 = 1–431; pVP16SAP90-5 = 69–150; pVP16SAP90-6 = 160–245. SAP102 pVP16SAP102-1 = 1–519. PSD-93 pVP16PSD-93-1 = 1–539. ZO1 pVP16ZO1-1 = 1–500. dlg-A pVP16dlg-1 = 1–598. Nitric oxide synthase pVP16NOS = 1–101.
-
Doyle D. A., et al., Cell 85, 1067 (1996).
(
10.1016/S0092-8674(00)81307-0
) / Cell by Doyle D. A. (1996) - Synthetic peptides were purified on a Kromasil 100 C 18 reversed-phase column (sequences: NMDAR2A SNRRVYKKMPSIESDV; NL1-1 LPHPHPHPHSHSTTRV; NL1-2 LPHPHPHPHSHST; control KFIEAGQYNSHLYGTSV). NMDAR2A and NL1-1 correspond to the COOH-terminus of NMDA2A and neuroligin 1 respectively. NL1-2 is identical with NL1-1 except it lacks the last three amino acids. The control peptide is composed of residues 597 to 613 from PSD-95/SAP90. Peptides were immobilized on a CM5 research grade sensor chip with the amine coupling kit (Pharmacia) equilibrated with 50 mM Hepes-NaOH (pH 8.0) containing 100 mM NaCl and superfused with GST fusion proteins at different concentrations (flow rate 20 μl/min). Binding activities (in resonance units) were measured as the difference between the baseline value determined 10 s before sample injection and the measurements taken at the indicated time points. All experiments were performed at 25°C. Data were analyzed with the BIA Evaluation program 2.1 (Pharmacia) [
- Joehnsson U., et al., BioTechniques 11, 520 (1991)]. / BioTechniques by Joehnsson U. (1991)
- IgG fusion proteins of the extracellular domains of neurexin 1β with and without an insert in splice site 4 and control IgG fusion protein were purified from transfected COS cells (4). Twelve rat brains were homogenized in 48 ml of 20 mM Hepes-NaOH (pH 8.0) 1% (w/v) NP-40 0.2 M NaCl 2 mM EGTA and phenylmethylsulfonyl fluoride (0.1 g/liter) in a glass Teflon homogenizer and centrifuged (100 000 g for 60 min at 4°C). The supernatant (60 ml) was adjected to 3.5 mM CaCl 2 . Supernatant (20 ml) (0.2 to 0.3 g of protein) was incubated overnight at 4°C with recombinant IgG fusion proteins immobilized on protein A-Sepharose. Beads were washed repeatedly by centrifugation in 20 mM Hepes-NaOH (pH 8.0) 2.5 mM CaCl 2 0.2 M NaCl and 0.5% NP-40 and eluted sequentially with 0.3 ml of the wash buffer containing 0.5 M NaCl and with 0.5 ml of 20 mM Hepes-NaOH (pH 8.0) 0.1 M NaCl 10 mM EGTA and 0.5% NP-40. Fractions were analyzed by SDS–polyacrylamide gel electrophoresis and by immunoblotting with antibodies to PSD-95 (polyclonal antiserum L667 and two independent monoclonal antibodies) and to neuroligin 1 (polyclonal antiserum L067). To exclude the possibility that PSD-95 directly interacts with neurexin 1β extracts from COS cells transfected with full-length PSD-95 were affinity chromatographed on immobilized neurexin 1β in the absence and presence of recombinant neuroligin.
- HEK293 cells were transfected with pCMVPSD95-1 encoding full-length rat PSD-95 either alone or together with pCMVNL18 pCMVNR2A or pCMVNR1 encoding full-length neuroligin 1 NMDA2A or NMDA1 respectively. Cells were stained with a mouse monoclonal antibody to PSD-95 and rabbit polyclonal antibodies to neuroligin 1 (L067) or the two NMDA receptors (from Chemicon). Images were obtained on a Bio-Rad MRC1024 confocal microscope.
10.1523/JNEUROSCI.16-08-02488.1996
- Russell A. B., Carlson S. S., ibid. 17, 4734 (1997). / ibid. by Russell A. B. (1997)
-
N. Uchida Y. Honjo K. R. Johnson M. J. Wheelock M. Takeichi J. Cell Biol. 135 767.
(
10.1083/jcb.135.3.767
) - Abbreviations for amino acids are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp; Y Tyr.
- We thank A. Roth E. Borowicz and I. Leznicki for excellent technical assistance; M. B. Kennedy (CalTech) for monoclonal antibodies to PSD-95; S. Nakanishi (Kyoto) for NMDA receptor cDNA clones; and M. Missler M. S. Brown and J. L. Goldstein for advice. Partially supported by grants from the NIH (RO1-MH52804) the Perot Family Foundation and the ERATO (Japan Science and Technology Co.). Y.H. was supported by a postdoctoral fellowship from the HFSP and T.W.R. from the DFG.
Dates
Type | When |
---|---|
Created | 23 years ago (July 27, 2002, 5:50 a.m.) |
Deposited | 1 year, 7 months ago (Jan. 12, 2024, 10:52 p.m.) |
Indexed | 1 day, 6 hours ago (Aug. 24, 2025, 7:08 p.m.) |
Issued | 27 years, 11 months ago (Sept. 5, 1997) |
Published | 27 years, 11 months ago (Sept. 5, 1997) |
Published Print | 27 years, 11 months ago (Sept. 5, 1997) |
@article{Irie_1997, title={Binding of Neuroligins to PSD-95}, volume={277}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.277.5331.1511}, DOI={10.1126/science.277.5331.1511}, number={5331}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Irie, M. and Hata, Y. and Takeuchi, M. and Ichtchenko, K. and Toyoda, A. and Hirao, K. and Takai, Y. and Rosahl, T. W. and Südhof, T. C.}, year={1997}, month=sep, pages={1511–1515} }