An Intramolecular Interaction between Src Homology 3 Domain and Guanylate Kinase-Like Domain Required for Channel Clustering by Postsynaptic Density-95/SAP90

Shin, Hye‐Won; Hsueh, Yi‐Ping; Yang, Fu-Chia; Kim, Eunjoon; Sheng, Morgan

doi:10.1523/jneurosci.20-10-03580.2000

Cited by 120 publications

(105 citation statements)

References 44 publications

Supporting

Mentioning

102

Contrasting

Order By: Relevance

“…From these observations we may assume that the "closed," cis-interacting conformation of the SH3-GK region (21,29,43) could prevent binding of ligands to the SH3 domain and/or to the GK domain. The binding of high affinity ligands such as CaM is thought to "open" the compact, intramolecularly stabilized SH3-HOOK-GK region and allow access of proteins to the GK domain (28). This view is in line with the inhibitory effect of the SH3 domain on the interaction of the GK domain with GKAP, which was already described (21), and it is reminiscent of the regulation mode of Src family tyrosine kinases (47).…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 81%

“…This leucine residue is conserved among MAGUK family members, and L612P corresponds to a point mutation that was first identified in the Drosophila dlg gene (dlg m30 allele; L632P mutation) where it produced a strong phenotype associated with loss of normal cell proliferation control (33). Moreover, the equivalent L460P mutation in the PSD-95 protein has been shown to interrupt intramolecular interaction between the SH3 and GK domains (27,28). The data we obtained for the SAP97(L612P) protein were very similar to those of the SH3-free protein (SAP97-2 construct), suggesting that calmodulin most efficiently binds to the intact protein in which intramolecular association of the SH3 and GK domains can occur.…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

“…CaM binding to SAP97 could have a similar function by relieving SH3 domain interference with the GK domain-GKAP interaction (21). In fact, modulation of the intramolecular SH3-GK interaction has been proposed as a general mechanism for regulating the clustering of the Kv1.4 potassium channel by SAP90 (28). However, the effect of the SH3 domain on the interaction of BEGAIN and GAKIN with the GK domain has not been evaluated yet.…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

“…Because the SH3 and the GK domains of MAGUKs show both intramolecular and intermolecular interactions (11,(27)(28)(29), CaM binding to these sites may modulate the scaffolding properties of SAP97 by affecting the equilibrium between intraand intermolecular assembly. In fact, a CaM-dependent clustering of N-methyl-D-aspartic acid receptors by heteromeric MAGUK protein complexes has been proposed (24), and a pivotal role has been attributed to SAP97 as a major constituent of a membrane-anchoring complex that contains Ca 2ϩ /calmodulindependent protein kinase II, actin, actinin, and protein 4.1 (30).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Formation of Complexes between Ca2+·Calmodulin and the Synapse-associated Protein SAP97 Requires the SH3 Domain-Guanylate Kinase Domain-connecting HOOK Region

Paarmann

Spangenberg

Lavie

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

Mammalian synapse-associated protein SAP97, a structural and functional homolog of Drosophila Dlg, is a membrane-associated guanylate kinase (MAGUK) that is present at pre-and postsynaptic sites as well as in epithelial cell-cell contact sites. It is a multidomain scaffolding protein that shares with other members of the MAGUK protein family a characteristic modular organization composed of three sequential protein interaction motifs known as PDZ domains, followed by an Src homology 3 (SH3) domain, and an enzymatically inactive guanylate kinase (GK)-like domain. Specific binding partners are known for each domain, and different modes of intramolecular interactions have been proposed that particularly involve the SH3 and GK domains and the so-called HOOK region located between these two domains. We identified the HOOK region as a specific site for calmodulin binding and studied the dynamics of complex formation of recombinant calmodulin and SAP97 by surface plasmon resonance spectroscopy. Binding of various SAP97 deletion constructs to immobilized calmodulin was strictly calciumdependent. From the rate constants of association and dissociation we determined an equilibrium dissociation constant K d of 122 nM for the association of calcium-saturated calmodulin and a SAP97 fragment, which encompassed the entire SH3-HOOK-GK module. Comparative structure-based sequence analysis of calmodulin binding regions from various target proteins predicts variable affinities for the interaction of calmodulin with members of the MAGUK protein family. Our findings suggest that calmodulin could regulate the intramolecular interaction between the SH3, HOOK, and GK domains of SAP97.Membrane-associated guanylate kinase homologs (MAGUKs) 1 have been implicated in the assembly of synapses and tight junctions. These are multidomain proteins consisting of one or more PDZ domains, an Src homology 3 (SH3) domain, and a guanylate kinase (GK)-like domain (for reviews, see Refs. 1-4). Rat SAP97 belongs to the MAGUK subfamily comprising Drosophila Dlg, SAP97/hDlg, SAP90/PSD-95, SAP102/NE-Dlg, and PSD93/Chapsyn110. SAP97 has earlier been reported to be presynaptic (5), but it has recently been shown also to be present at postsynaptic sites in cerebral cortex (6); it is found both at the post-synaptic density (PSD) region and in the cytoplasm of hippocampal synapses, suggesting a role for SAP97 in ionotropic glutamate receptor trafficking (7). Recently it was discovered that SAP97, via its guanylate kinase (GK)-like domain, directly regulates the function of an inwardly rectifier potassium channel (8). In addition to its crucial role as a scaffolding protein in neuronal cells, SAP97 is an essential component of the basolateral membrane cytoskeleton in a variety of epithelial cells (5).Each domain in the SAP subfamily of MAGUKs is highly conserved and has been shown to be a site of protein-protein interaction: PDZ domains bind voltage-and ligand-gated ion channels (3, 9); SH3 domains interact with proline-rich, PXXPR-like sequences (3, 4, 10) as w...

show abstract

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 81%

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Formation of Complexes between Ca2+·Calmodulin and the Synapse-associated Protein SAP97 Requires the SH3 Domain-Guanylate Kinase Domain-connecting HOOK Region

Paarmann

Spangenberg

Lavie

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…However, interaction in the neurons may be regulated by phosphorylation-dependent conformational changes to the full-length proteins. For example, the SH3 and GK domains of PSD-95 form an intramolecular association that regulates coclustering of PSD-95 with ion channels in heterologous cells (McGee and Bredt, 1999;Shin et al, 2000). Thus PKA phosphorylation may determine the availability of a key protein-protein interaction domain.…”

Section: Mechanisms Of Activity-regulated Synaptic Targeting Of Nmda mentioning

confidence: 99%

cAMP-Dependent Protein Kinase Mediates Activity-Regulated Synaptic Targeting of NMDA Receptors

2001

View full text Add to dashboard Cite

Chronic activity blockade increases synaptic levels of NMDA receptor immunoreactivity in hippocampal neurons. We show here that blockade-induced synaptic NMDA receptors are functional and mediate enhanced excitotoxicity in response to synaptically released glutamate. Activity blockade increased the cell surface association of NMDA receptors. Blockade-induced synaptic targeting of NMDA receptors did not require protein synthesis but required phosphorylation and specifically cAMPdependent protein kinase (PKA). Furthermore, activation of PKA was sufficient to induce synaptic targeting of NMDA receptors regardless of receptor activity status. These results implicate PKA activity downstream of receptor blockade as a mediator of enhanced synaptic transport or stabilization of NMDA receptors. Synaptic clustering of NR1-green fluorescent protein was observed in living neurons in response to NMDA receptor and cAMP phosphodiesterase antagonists and occurred gradually over the course of a day. This pathway represents a cellular mechanism for synaptic homeostasis and is likely to function in metaplasticity, long-term regulation of the ability of a synapse to undergo potentiation or depression. Key words: NMDA receptor; synaptogenesis; activity; synaptic clustering; excitotoxicity; subcellular localization; hippocampus; NR1-GFPThe NMDA-type glutamate receptor plays a central role in circuit development, memory formation, and many forms of synaptic plasticity in the mammalian brain. The NMDA receptor is composed of the essential NR1 subunit and one or more of the modulatory NR2A-D and NR3 subunits (Nakanishi, 1992;Seeburg, 1993;Mori and Mishina, 1995). NMDA receptor channel opening requires ligand binding (by presynaptic glutamate release) and removal of Mg 2ϩ block (by postsynaptic depolarization), thus conferring on the NMDA receptor the ability to function as a molecular coincidence detector (Mayer et al., 1984). Through its Ca 2ϩ permeability, NMDA receptor function is linked with many downstream signal transducing pathways in the neuron. The magnitude and kinetics of calcium elevation at the synapse are thought to be major determinants of long-term effects on synaptic efficacy (Lisman, 1989;Abraham and Bear, 1996).The level of NMDA receptor function at the synapse critically regulates brain function and cell survival. Mice expressing 5% of normal levels of NR1 exhibit increased motor activity, stereotypy, and deficits in social and sexual interactions, behaviors associated with schizophrenia (Mohn et al., 1999). Deletion of NR1 targeted postnatally selectively to CA1 of the hippocampus results in mice that are viable but deficient in spatial learning and formation of temporal memory (Tsien et al., 1996;Huerta et al., 2000). In contrast, overactivation of NMDA receptors contributes substantially to neuronal death during epilepsy, stroke, trauma, and neurodegenerative disorders (McDonald and Johnston, 1990;Choi, 1994;Rothman and Olney, 1995;During et al., 2000).NMDA receptor function is regulated during development and by ...

show abstract

Role of the maguk protein family in synapse formation and function

Escobedo

Astorga

Molina

et al. 2011

Developmental Neurobiology

100

View full text Add to dashboard Cite

Synaptic function is crucially dependent on the spatial organization of the presynaptic and postsynaptic apparatuses and the juxtaposition of both membrane compartments. This precise arrangement is achieved by a protein network at the submembrane region of each cell that is built around scaffold proteins. The membrane-associated guanylate kinase (MAGUK) family of proteins is a widely expressed and well-conserved group of proteins that plays an essential role in the formation and regulation of this scaffolding. Here, we review general features of this protein family, focusing on the discs large and calcium/calmodulin-dependent serine protein kinase subfamilies of MAGUKs in the formation, function, and plasticity of synapses.

show abstract

An Intramolecular Interaction between Src Homology 3 Domain and Guanylate Kinase-Like Domain Required for Channel Clustering by Postsynaptic Density-95/SAP90

Cited by 120 publications

References 44 publications

Formation of Complexes between Ca2+·Calmodulin and the Synapse-associated Protein SAP97 Requires the SH3 Domain-Guanylate Kinase Domain-connecting HOOK Region

Formation of Complexes between Ca2+·Calmodulin and the Synapse-associated Protein SAP97 Requires the SH3 Domain-Guanylate Kinase Domain-connecting HOOK Region

cAMP-Dependent Protein Kinase Mediates Activity-Regulated Synaptic Targeting of NMDA Receptors

Role of the maguk protein family in synapse formation and function

Contact Info

Product

Resources

About