Interaction of the Nav1.2a Subunit of the Voltage-dependent Sodium Channel with Nodal AnkyrinG

Bouzidi, Mourad; Tricaud, Nicolas; Giraud, Pierre; Kordeli, Ekaterini; Caillol, Ghislaine; Deleuze, Charlotte; Couraud, François; Alcaraz, Gisèle

doi:10.1074/jbc.m201760200

Cited by 53 publications

(32 citation statements)

References 50 publications

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“…Further studies in which individual cytoplasmic loops of the III-IV construct were replaced sequentially with the nonbinding N terminus of rNa v 1.2a confirmed that the binding site was restricted to loop II-III (not shown). These results are in contrast to the findings of Bouzidi et al (18) who recently reported interactions between ankyrin G and fusion proteins representing the isolated intracellular loops between domains I-II and III-IV but not between the II-III loop and ankyrin G .…”

Section: Na V 12 Co-localizes With Ankyrin G In Vivo and Binds To Itscontrasting

confidence: 56%

“…Regulated interactions between ankyrin and VGSC-␤ subunits have been detected using a cell-based recruitment assay (17,21). In contrast to these results, Bouzidi et al (18) did not detect interactions between ankyrin G and the cytoplasmic domains of the ␤ subunits using bacterially expressed proteins in an in vitro binding assay. However, they did detect interactions with recombinant fusion proteins representing the I-II (loop 1) and the III-IV (loop 3) cytoplasmic linkers.…”

Section: Discussionmentioning

confidence: 64%

“…Initial studies using uncharacterized rat brain preparations of VGSCs and purified erythrocyte ankyrin (15) did not distinguish which VGSC subunits or their isoforms were involved in ankyrin binding. Subsequent studies using a variety of techniques have suggested sites for ankyrin interaction on both the VGSC ␤ (17) and ␣ subunits (18).…”

mentioning

confidence: 99%

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Identification of a Conserved Ankyrin-binding Motif in the Family of Sodium Channel α Subunits

Lemaillet

Walker

Lambert

2003

Journal of Biological Chemistry

226

231

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Interactions with ankyrin G are crucial to the localization of voltage-gated sodium channels (VGSCs) at the axon initial segment and for neurons to initiate action potentials. However, the molecular nature of these interactions remains unclear. Here we report that VGSC-␣, but not -␤, subunits bind to ankyrin G using pull-down assays. Further dissection of this activity identifies a conserved 9-amino acid motif ((V/A)P(I/L)AXXE(S/D)D) required for ankyrin G binding. This motif is also required for the localization of chimeric neurofascin/sodium channel molecules to the initial segment of cultured hippocampal neurons. The conserved nature of this motif suggests that it functions to localize sodium channels to a variety of "excitable" membrane domains both inside and outside of the nervous system.The concentration of voltage-gated sodium channels (VGSCs) 1 into excitable membrane domains is crucial to information processing and transmission in the nervous system and to excitation/contraction coupling in muscle. Localized concentrations of VGSCs at the initial segment (IS) and the nodes of Ranvier are necessary for the initiation and propagation of action potentials through myelinated axons. However, mechanisms underlying the localization of VGSCs to these excitable membrane domains remain relatively unknown. In neurons, VGSCs exist as heterotrimers composed of a large pore-forming ␣ subunit associated with two smaller accessory ␤ subunits (reviewed in Ref. 1). At least 10 genes encoding putative ␣ subunits have been identified in mammals (reviewed in Ref.2) along with three ␤ subunit genes. ␤ subunits not only modulate the activity of ␣ subunits (3, 4) but also exhibit characteristics of cell adhesion molecules (CAMs) binding to extracellular matrix molecules such as tenascin-C and -R (5) and other CAMs such as neurofascin (6).Polarized localization of VGSCs in the axonal membrane requires interactions with members of the ankyrin family of peripheral membrane proteins (reviewed in Ref. 7). Ankyrins function as membrane-cytoskeleton adaptors that immobilize integral membrane proteins to the spectrin-based membrane skeleton. Ankyrins co-purify with and directly bind to purified VGSCs (8), and specific isoforms of the ankyrin G gene are concentrated with VGSCs at the IS (9), the nodes of Ranvier (9), and the neuromuscular junction (10). The functional importance of the interaction between ankyrin G and VGSCs is demonstrated in knockouts of ankyrin G in the mouse cerebellum. Purkinje cells from knockout animals are deficient in localized VGSC concentrations at the IS (11) and are unable to initiate action potentials (12).In addition to VGSCs, ankyrins also interact with a variety of other integral membrane proteins present at the node including the CAMs neurofascin and NrCAM (13,14). Interactions of ankyrins with VGSCs (15) and other integral membrane proteins are mediated through the N-terminal 90-kDa repeat or membrane-binding (MB) domain. The interaction between the ankyrin MB domain and neurofascin/NrCAM is depende...

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Section: Na V 12 Co-localizes With Ankyrin G In Vivo and Binds To Itscontrasting

confidence: 56%

Section: Discussionmentioning

confidence: 64%

See 1 more Smart Citation

Identification of a Conserved Ankyrin-binding Motif in the Family of Sodium Channel α Subunits

Lemaillet

Walker

Lambert

2003

Journal of Biological Chemistry

226

231

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“…For example, ankyrin, which is a family of ubiquitous adapter proteins linking membrane proteins to the spectrin-actin cytoskeleton, is a well known major interacting protein of Na v 1.2 (Jenkins and Bennett, 2002;Boiko et al, 2003). For Na v 1.2, the intracellular linker between domains I and II together with the N-terminal region has been proposed as a major interaction site for ankyrin (Bouzidi et al, 2002). Furthermore, the cytoskeleton has been known to modulate sodium channel gating (Matsumoto and Sakai, 1979a,b;Fukuda et al, 1981).…”

Section: Discussionmentioning

confidence: 99%

A Nonsense Mutation of the Sodium Channel GeneSCN2Ain a Patient with Intractable Epilepsy and Mental Decline

Kamiya¹,

Kaneda²,

Sugawara³

et al. 2004

J. Neurosci.

186

146

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Mutations, exclusively missense, of voltage-gated sodium channel ␣ subunit type 1 (SCN1A) and type 2 (SCN2A) genes were reported in patients with idiopathic epilepsy: generalized epilepsy with febrile seizures plus. Nonsense and frameshift mutations of SCN1A, by contrast, were identified in intractable epilepsy: severe myoclonic epilepsy in infancy (SMEI). Here we describe a first nonsense mutation of SCN2A in a patient with intractable epilepsy and severe mental decline. The phenotype is similar to SMEI but distinct because of partial epilepsy, delayed onset (1 year 7 months), and absence of temperature sensitivity. A mutational analysis revealed that the patient had a heterozygous de novo nonsense mutation R102X of SCN2A. Patch-clamp analysis of Na v 1.2 wild-type channels and the R102X mutant protein coexpressed in human embryonic kidney 293 cells showed that the truncated mutant protein shifted the voltage dependence of inactivation of wild-type channels in the hyperpolarizing direction. Analysis of the subcellular localization of R102X truncated protein suggested that its dominant negative effect could arise from direct or indirect cytoskeletal interactions of the mutant protein. Haploinsufficiency of Na v 1.2 protein is one plausible explanation for the pathology of this patient; however, our biophysical findings suggest that the R102X truncated protein exerts a dominant negative effect leading to the patient's intractable epilepsy.

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“…This suggests that ankyrinG and Na V 1 bind directly to each other at the earliest stage of Na V 1 channel clustering. Biochemical evidence exists for an interaction in vitro between the N-terminal end of ankyrin and Na V 1 (Srinivasan et al, 1992;Bouzidi et al, Teased fiber preparations allow NMJs to be viewed en face. In the region of high AChR density, the density of labeling for Na V 1 (B) and ␣-fodrin is low ( C), whereas that for utrophin is high 2002).…”

Section: Discussionmentioning

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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Interaction of the Nav1.2a Subunit of the Voltage-dependent Sodium Channel with Nodal AnkyrinG

Cited by 53 publications

References 50 publications

Identification of a Conserved Ankyrin-binding Motif in the Family of Sodium Channel α Subunits

Identification of a Conserved Ankyrin-binding Motif in the Family of Sodium Channel α Subunits

A Nonsense Mutation of the Sodium Channel GeneSCN2Ain a Patient with Intractable Epilepsy and Mental Decline

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

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