Specific interactions between the syntrophin PDZ domain and voltage-gated sodium channels

Schultz, Johan; Hoffmuüller, Ulrich; Krause, Gerd; Ashurst, Jennifer; Macias, María J.; Schmieder, Peter; Schneider‐Mergener, Jens; Oschkinat, Hartmut

doi:10.1038/nsb0198-19

Cited by 216 publications

(215 citation statements)

References 32 publications

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“…In PSD-95 PDZ3, Arg-318 coordinates a water molecule that in turn stabilizes the negative charge on the bound carboxylate, together with the backbone amides (11). Harris et al (14) concluded that Lys-86 in ␣1-syntrophin PDZ is not involved in a direct ionic interaction with the peptide carboxylate, in agreement with the published structures (11,12,23,26,27) as well as our data. The equilibrium constant of ␣1-syntrophin PDZ domain binding to a C-terminal peptide is shown to display a salt dependence, with lower affinity at higher salt, similarly as PDZ3 in the present study (14).…”

Section: Discussionsupporting

confidence: 92%

“…Arg-318 is linked to the carboxylate of the peptide C-terminal valine via an ordered water molecule (11). Glu-373 of the ␣B helix corresponds to Asp-143 of the mouse ␣1-syntrophin PDZ that is known to form a salt bridge with a lysine residue at position Ϫ4 of its peptide (23). Kinetic experiments similar to those for the wild type were done on the mutants (Table 2).…”

Section: Resultsmentioning

confidence: 99%

“…The negatively charged C-terminal carboxylate is bound via hydrogen bonds to the protein backbone NH groups of the so-called carboxylate-binding loop (11,12,23,26,27), (Fig. 1b).…”

Section: Discussionmentioning

confidence: 99%

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Two Conserved Residues Govern the Salt and pH Dependencies of the Binding Reaction of a PDZ Domain

Celestine

Engström

Gianni

et al. 2006

Journal of Biological Chemistry

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PDZ domains are protein-protein interaction modules found in hundreds of human proteins. Their binding reactions are sensitive to variations in salt and pH but the basis of the respective dependence has not been clear. We investigated the binding reaction between PSD-95 PDZ3 and a peptide corresponding to a native ligand with protein engineering in conjunction with stopped-flow and equilibrium fluorimetry and found that the two conserved residues Arg-318 and His-372 were responsible for the salt and pH dependencies, respectively. The basis of the salt-dependent variation of the affinity was explored by mutating all charged residues in and around the peptide-binding pocket. Arg-318 was found to be crucial, as mutation to alanine obliterated the effect of chloride on the binding constants. The direct interaction of chloride with Arg-318 was demonstrated by time-resolved urea denaturation experiments, where the Arg-318 3 Ala mutant was less stabilized by addition of chloride as compared with wild-type PDZ3. We also demonstrated that protonation of His-372 was responsible for the increase of the equilibrium dissociation constant at low pH. Both chloride concentration and pH (during ischemia) vary in the postsynaptic density, where PSD-95 is present, and the physiological buffer conditions may thus modulate the interaction between PSD-95 and its ligands through binding of chloride and protons to the "molecular switches" Arg-318 and His-372, respectively.In the post-genomic era there will be ever-increasing focus on protein-protein interactions (1-3) because of their critical role in cellular function and the growing sequence and structure databases. Indeed, most processes in the cell are governed by more or less specific protein-protein interactions. Often the binding is tuned by cellular or extracellular conditions such as pH, ionic strength, and molecular crowding, and also by specific molecules acting allosterically on the protein or competing with the "natural" ligand. Many proteins or protein domains, have evolved to interact with other proteins and form complexes, and one of the most common of these protein-protein interaction modules is the PDZ domain, which is present in several hundred human proteins (4, 5). PDZ domains are often found within multi-domain scaffolding proteins, and they bind mainly to the C termini of their target proteins (6 -8) but also internally (9, 10). PDZ domains have been shown to be rather promiscuous with regard to their ligand, and they have overlapping ligand specificities (6, 7).One of the most well studied PDZ domains is PDZ3 from PSD-95.3 The three-dimensional structure of this protein with and without its target ligand has been solved (11). The canonical PDZ domain consists of about 90 -100 amino acids forming six ␤ strands (␤A to ␤F) and two ␣ helices; ␣A and ␣B neatly arranged in a spherical structure (Fig. 1a). Peptide binding takes place between the ␤B strand and ␣B helix in an antiparallel manner as shown in Fig. 1a. As judged by the crystal structure, PDZ3 binds its targ...

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Section: Discussionsupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Two Conserved Residues Govern the Salt and pH Dependencies of the Binding Reaction of a PDZ Domain

Celestine

Engström

Gianni

et al. 2006

Journal of Biological Chemistry

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“…Although syntrophins have been shown to bind directly to muscle Na V 1s (Gee et al, 1998;Schultz et al, 1998), their role in concentrating Na V 1s at the NMJ remains unclear. Interactions with syntrophin are not required for the association of Na V 1 with the plasma membrane (Adams et al, 2001).…”

Section: Stabilization Of Na V 1 Clusters At Maturing Nmjsmentioning

confidence: 99%

Voltage-Gated Sodium Channels and AnkyrinG Occupy a Different Postsynaptic Domain from Acetylcholine Receptors from an Early Stage of Neuromuscular Junction Maturation in Rats

et al. 2003

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Spatial segregation of membrane proteins is a feature of many excitable cells. In skeletal muscle, clusters of acetylcholine receptors (AChRs) and voltage-gated sodium channels (Na V 1s) occupy distinct domains at the neuromuscular junction (NMJ). We used quantitative immunolabeling of developing rat soleus muscles to study the mechanism of ion channel segregation and Na V 1 clustering at NMJs. When Na V 1s can first be detected, at birth, they already occupy a postsynaptic domain that is distinct from that occupied by AChRs. At this time, Na V 1s are expressed only in a diffuse area that extends 50 -100 m from the immature NMJ. However, in the region of the high-density AChR cluster at NMJ itself, Na V 1s are actually present in lower density than in the immediately surrounding membrane. These distinctive features of the Na V 1 distribution at birth are closely correlated with the distribution of ankyrinG immunolabeling. This suggests that an interaction with ankyrinG plays a role in the initial segregation of Na V 1s from AChRs. Both Na V 1 and ankyrinG become clustered at the NMJ itself 1-2 weeks after birth, coincident with the formation of postsynaptic folds. Syntrophin immunolabeling codistributes with AChRs and never resembles that for Na V 1 or ankyrinG. Therefore, syntrophin is unlikely to play an important part in the initial accumulation of Na V 1 at the NMJ. These findings suggest that the segregation of Na V 1 from AChRs begins early in NMJ formation and occurs as a result of the physical exclusion of Na V 1 and ankyrinG from the region of nerve-muscle contact rather than by a process of active clustering.

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“…Binding studies show that both the first and second PDZ repeats in PSD-95 potently interact with the COOH-terminal tails of specific NMDA receptor subunits and other ion channels that terminate in a consensus Glu-(Ser/Thr)-X-(Val/Ile)-COOH (15)(16)(17)(18). Crystallographic studies have determined the structural design of this PDZ-peptide interface, which requires: 1) sequencespecific interactions and 2) peptide termination immediately following the valine (19,20). The fact that PSD-95 PDZ2 can interact with both COOH-terminal peptides and with the nNOS PDZ domain is intriguing.…”

mentioning

confidence: 99%

PSD-95 Assembles a Ternary Complex with theN-Methyl-d-aspartic Acid Receptor and a Bivalent Neuronal NO Synthase PDZ Domain

Christopherson

Hillier

Lim

et al. 1999

Journal of Biological Chemistry

514

362

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Nitric oxide (NO) biosynthesis in cerebellum is preferentially activated by calcium influx through N-methyl-D-aspartate (NMDA)-type glutamate receptors, suggesting that there is a specific link between these receptors and neuronal NO synthase (nNOS). Here, we find that PSD-95 assembles a postsynaptic protein complex containing nNOS and NMDA receptors. Formation of this complex is mediated by the PDZ domains of PSD-95, which bind to the COOH termini of specific NMDA receptor subunits. In contrast, nNOS is recruited to this complex by a novel PDZ-PDZ interaction in which PSD-95 recognizes an internal motif adjacent to the consensus nNOS PDZ domain. This internal motif is a structured "pseudo-peptide" extension of the nNOS PDZ that interacts with the peptide-binding pocket of PSD-95 PDZ2. This asymmetric interaction leaves the peptidebinding pocket of the nNOS PDZ domain available to interact with additional COOH-terminal PDZ ligands. Accordingly, we find that the nNOS PDZ domain can bind PSD-95 PDZ2 and a COOH-terminal peptide simultaneously. This bivalent nature of the nNOS PDZ domain further expands the scope for assembly of protein networks by PDZ domains.Efficiency and specificity in cellular signaling cascades is often mediated by assembly of multiprotein transduction networks. Signaling by the calcium/calmodulin regulated neuronal nitric oxide synthase (nNOS) 1 in cerebellar neurons is activated by calcium influx through N-methyl-D-aspartic acid (NMDA) receptors (1, 2), but nNOS is not efficiently stimulated by activation of non-NMDA receptors that also generate calcium influx (3). Therefore, a mechanism must exist to specifically couple NMDA receptor-mediated calcium influx to nNOS.In addition to mediating neuronal functions, nNOS is also found in skeletal muscle, where nNOS localizes to the sarcolemma. Membrane association of nNOS in skeletal muscle is mediated by direct interaction with ␣1-syntrophin, a component of the dystrophin complex. Binding of nNOS to syntrophin occurs through direct interaction of PDZ protein motifs present near the NH 2 termini of both proteins (4). Mice lacking ␣1-syntrophin lose nNOS protein and enzyme activity from muscle membranes (5). Furthermore, patients with Duchenne muscular dystrophy and mdx mice that lack dystrophin evince a selective loss of nNOS from the sarcolemma (6).In neurons, nNOS is also associated with cell membranes. In a detailed ultrastructural analysis of primate visual cortex, nNOS immunoreactivity in spines was concentrated over thick postsynaptic specializations of plasma membranes, often in association with NMDA receptors (7). This synaptic localization of nNOS in brain may be mediated by association with the postsynaptic density protein, PSD-95. Like ␣1-syntrophin, PSD-95 binds directly to nNOS through a PDZ-PDZ interaction that involves the second PDZ domain of PSD-95 (4). Moreover, PSD-95 is a member of a larger family of postsynaptic density proteins (PSD-95/SAP-90, PSD-93/chapsyn-110, and SAP-102) that cluster NMDA receptors and may anchor these...

show abstract

Specific interactions between the syntrophin PDZ domain and voltage-gated sodium channels

Cited by 216 publications

References 32 publications

Two Conserved Residues Govern the Salt and pH Dependencies of the Binding Reaction of a PDZ Domain

Two Conserved Residues Govern the Salt and pH Dependencies of the Binding Reaction of a PDZ Domain

Voltage-Gated Sodium Channels and AnkyrinG Occupy a Different Postsynaptic Domain from Acetylcholine Receptors from an Early Stage of Neuromuscular Junction Maturation in Rats

PSD-95 Assembles a Ternary Complex with theN-Methyl-d-aspartic Acid Receptor and a Bivalent Neuronal NO Synthase PDZ Domain

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