The N -methyl-D-aspartate (NMDA) receptor subserves synaptic glutamate-induced transmission and plasticity in central neurons. yeast two-hybrid system was used to show that the cytoplasmic tails of NMDA subunits interact with a prominent postsynaptic density protein PSD-95. second PDZ domain PSD-95 binds seven-amino acid, COOH-terminal containing terminal t SXV motif (where S is serine, X any amino V valine) common NR2 certain NR1 splice forms. Transcripts encoding are expressed pattern...

By three criteria, two biochemical and one immunochemical, the major postsynaptic density protein (mPSDp) is indistinguishable from 50-kilodalton (kDa) alpha subunit of a brain calmodulin-dependent kinase. First, proteins comigrate on NaDodSO4/polyacrylamide gels. Second, iodinated tryptic peptide maps are identical. Finally, monoclonal antibody (6G9) that was raised against kinase binds immunoblots to single 50 kDa band in crude homogenates both purified mPSDp fractions. The holoenzyme also...

A calcium and calmodulin-dependent protein kinase has been purified from rat brain. It was monitored during the purification by its ability to phosphorylate synaptic vesicle-associated protein, synapsin I. 300-fold sufficient produce that is 90-95% pure as determined scans of stained sodium dodecyl sulfate-polyacrylamide gels a specific activity 2.9 mumol 32P transferred per min/mg protein. Thus, relatively abundant brain enzyme, perhaps comprising much 0.3% total The Stokes radius (95 A)...

Two calcium-stimulated protein kinase activities (ATP:protein phosphotransferase, EC 2.7.1.37) that phosphorylate I, a specific synaptic protein, have been identified in homogenates of rat brain. One these is found both the particulate and cytosolic fractions phosphorylates region I phosphorylated intact synaptosomes response to calcium but not cyclic AMP. The stimulation by enzyme partially purified requires addition calmodulin. It yet known whether enzymes are related. A second only...

Our understanding of the organization postsynaptic signaling systems at excitatory synapses has been aided by identification proteins in density (PSD) fraction, a subcellular fraction enriched structures with morphology PSDs. In this study, we have completed most major PSD use an analytical method based on mass spectrometry coupled searching protein sequence databases. At least one each 26 prominent bands from now identified. We found 7 not previously known to be constituents and 24 that had...

The postsynaptic density (PSD) is a specialization of the submembranous cytoskeleton that visible in electron microscope on cytoplasmic face membrane. A subcellular fraction enriched structures with morphology PSDs contains signal-transduction molecules thought to regulate receptor localization and function central nervous system. We have purified prominent tyrosine-phosphorylated glycoprotein apparent molecular mass 180 kDa, termed PSD-gp180, highly rat forebrain PSD fraction. sequences...

The N-methyl-D-aspartate (NMDA) subtype of excitatory glutamate receptors plays critical roles in embryonic and adult synaptic plasticity the central nervous system. receptor is a heteromultimer core subunits, NR1, one or more regulatory NR2A-D. Protein phosphorylation can regulate NMDA function (Lieberman, D. N., Mody, I. (1994) Nature 369, 235-239; Wang, Y. T., Salter, M. W. 233-235; L.-Y., Orser, B. A., Brautigan, L., MacDonald, J. F. 230-232). Here we identify major site on subunit NR2B...

Forebrain and cerebellar Type I1 Ca2+/calmodulindependent protein kinases have different subunit compositions.The forebrain holoenzyme, characterized in our laboratory, is a 650-kDa holoenzyme composed of 50-kDa a-subunits 60-kDa &subunits assembled approximately 3:l ratio (Bennett, M. K., Erondu,

mRNA for the α-subunit of CaMKII is abundant in dendrites neurons forebrain (Steward, 1997). Here we show that tetanic stimulation Schaffer collateral pathway causes an increase concentration α-CaMKII postsynaptic neurons. The blocked by anisomycin and detected both quantitative immunoblot semiquantitative immunocytochemistry. dendritic can be measured 100–200 μm away from neuronal cell bodies as early 5 min after a tetanus. Transport mechanisms macromolecules are not fast enough to account...

cDNA clones coding for the beta subunit of rat brain type II Ca2+/calmodulin-dependent protein kinase were isolated and sequenced. The clones, including one containing entire region, hybridize at high stringency to a single band poly(A)+ RNA length 4.8 kilobases. coded by was identified in vitro transcription followed translation resulting RNA. DNA sequence contains long open reading frame (1626 nucleotides) 542 amino acids with molecular weight 60,333, amino-terminal half which is...

Autophosphorylation of calcium/calmodulin-dependent protein kinase II (CaMKII) at threonine-286 produces Ca 2+ -independent activity and has been proposed to be involved in induction long-term potentiation by tetanic stimulation the hippocampus. We have used an immunocytochemical method visualize quantify pattern autophosphorylation CaMKII hippocampal slices after tetanization Schaffer collateral pathway. Thirty minutes stimulation, autophosphorylated CaM (P-CaMKII) is significantly...

SynGAP is a brain-specific ras GTPase-activating protein that an abundant component of the signaling complex associated with NMDA-type glutamate receptor. We generated mutant mice lacking synGAP to study its physiological role. Homozygous die in first few days after birth; however, neurons from embryos can be maintained culture. Here, we report spine and synapse formation are accelerated cultured neurons, spines mature significantly larger than those wild type. Clusters PSD-95 subunits...