Structure of the SH3-Guanylate Kinase Module from PSD-95 Suggests a Mechanism for Regulated Assembly of MAGUK Scaffolding Proteins

McGee, Aaron W.; Dakoji, Srikanth; Olsen, Olav; Bredt, David S.; Lim, Wendell A.; Prehoda, Kenneth E.

doi:10.1016/s1097-2765(01)00411-7

Cited by 227 publications

(247 citation statements)

References 75 publications

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“…In both of the complex structures, the Dlg4 GK domain adopts a conformation highly similar to that from the Dlg1/Dlg4 SH3-GK tandem structures solved earlier, comprising the GMP-binding subdomain, the LID subdomain and the Core subdomain [20,23,24] (Figure 3A, 3B and 3E). Both of the p-Lgl2a and p-Lgl2c peptides adopt a short α-helical conformation in the complex, occupying the concave groove clamped by the GMPbinding subdomain (β3, β4, β5 and β6) and the Core subdomain (α1 helix) with the same orientation.…”

Section: Overall Structures Of the Dlg4 Gk/p-lgl2 Complexessupporting

confidence: 56%

Phosphorylation-dependent interaction between tumor suppressors Dlg and Lgl

et al. 2014

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Section: Overall Structures Of the Dlg4 Gk/p-lgl2 Complexessupporting

confidence: 56%

Phosphorylation-dependent interaction between tumor suppressors Dlg and Lgl

et al. 2014

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“…The U1 region mutant V37G shows reduced binding with the polarity protein Par6 (Wang et al, 2004), the L27N domain mutant disrupts the interaction between PALS1 and PATJ (Roh et al, 2002), and the PALS1 L27C domain mutant does not bind Lin-7 (Kamberov et al, 2000). It has been shown that extensive intramolecular interactions exist between the C-terminal SH3 domain and the GUK domain of MAGUK proteins (McGee et al, 2001;Tavares et al, 2001). To explore the function of the C terminus of PALS1, which has been largely unknown, we generated a point mutation L379P in the SH3 domain that disrupts the intramolecular interaction between the SH3 domain and the GUK domain (Woods et al, 1996;Wu et al, 2000), and a truncation mutant delC (PALS1 1-345) that lacks both domains.…”

Section: Wild-type Pals1 and Two Pals1 Mutants Rescue The Defects In mentioning

confidence: 99%

PALS1 Regulates E-Cadherin Trafficking in Mammalian Epithelial Cells

Wang¹,

Chen

Margolis

2007

MBoC

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Protein Associated with Lin Seven 1 (PALS1) is an evolutionarily conserved scaffold protein that targets to the tight junction in mammalian epithelia. Prior work in our laboratory demonstrated that the knockdown of PALS1 in Madin Darby canine kidney cells leads to tight junction and polarity defects. We have created new PALS1 stable knockdown cell lines with more profound reduction of PALS1 expression, and a more severe defect in tight junction formation was observed. Unexpectedly, we also observed a severe adherens junction defect, and both defects were corrected when PALS1 wild type and certain PALS1 mutants were expressed in the knockdown cells. We found that the adherens junction structural component E-cadherin was not effectively delivered to the cell surface in the PALS1 knockdown cells, and E-cadherin puncta accumulated in the cell periphery. The exocyst complex was also found to be mislocalized in PALS1 knockdown cells, potentially explaining why E-cadherin trafficking is disrupted. Our results suggest a broad and evolutionarily conserved role for the tight junction protein PALS1 in the biogenesis of adherens junction.

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“…The interactions between the SAP97/hDlg multidomain protein, a member of the MAGUK protein family, and its putative binding partners have so far only been studied by qualitative or at best semi-quantitative cell biological methods using the yeast two-hybrid approach or co-immunoprecipitation techniques for detecting protein-protein association (21,42). Detailed studies of tertiary structures of MAGUKs have been confined to the C-terminal half of SAP90/PSD95 encompassing the SH3, HOOK, and GK domains (29,43), and our most recent work on human CASK has revealed distinctive structural features of the CASK GK binding module in comparison to enzymatically active guanylate kinase (19). However, no complex between a MAGUK and any interacting protein has been crystallized so far.…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

“…Second, the presence of the SH3 domain significantly increases the affinity. 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).…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

“…Role of the C-terminal Tail-At present, no quantitative data are available on the postulated dynamic equilibrium between open and closed conformations in SAP97 (21) and SAP90 (43). Remarkably, the last 12 residues C-terminal to the GK domain of SAP90 appear to play a critical role in forming the native conformation of the molecule (29), because they were shown to fold into a ␤-strand that actually is a structural component of the SH3 domain formed through three-dimensional domain swapping (43).…”

Section: Table I Characteristics Of Cam Interaction With Sap97 Constrmentioning

confidence: 99%

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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

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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...

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Structure of the SH3-Guanylate Kinase Module from PSD-95 Suggests a Mechanism for Regulated Assembly of MAGUK Scaffolding Proteins

Cited by 227 publications

References 75 publications

Phosphorylation-dependent interaction between tumor suppressors Dlg and Lgl

Phosphorylation-dependent interaction between tumor suppressors Dlg and Lgl

PALS1 Regulates E-Cadherin Trafficking in Mammalian Epithelial Cells

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

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