β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Nav1.7 in HEK293 cells

Laedermann, Cédric J.; Syam, Ninda; Pertin, Marie; Décosterd, Isabelle; Abriel, Hugues

doi:10.3389/fncel.2013.00137

Cited by 61 publications

(55 citation statements)

References 47 publications

(55 reference statements)

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“…It should be noted that coexpression versus association of α-and β-subunits per se are not synonymous; thus, for example, Na V β1 coexpression can affect the glycosylation of the α-subunit (27), and the glycosylation state of the α-subunit, rather than its physical association with a β-subunit, could be responsible for the altered pharmacology of the VGSC.…”

Section: Discussionmentioning

confidence: 99%

A disulfide tether stabilizes the block of sodium channels by the conotoxin μO§-GVIIJ

Gajewiak¹,

Azam²,

Imperial³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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A cone snail venom peptide, μO §-conotoxin GVIIJ from Conus geographus, has a unique posttranslational modification, S-cysteinylated cysteine, which makes possible formation of a covalent tether of peptide to its target Na channels at a distinct ligandbinding site. μO §-conotoxin GVIIJ is a 35-aa peptide, with 7 cysteine residues; six of the cysteines form 3 disulfide cross-links, and one (Cys24) is S-cysteinylated. Due to limited availability of native GVIIJ, we primarily used a synthetic analog whose Cys24 was S-glutathionylated (abbreviated GVIIJ SSG ). The peptide-channel complex is stabilized by a disulfide tether between Cys24 of the peptide and Cys910 of rat (r) Na V 1.2. A mutant channel of rNa V 1.2 lacking a cysteine near the pore loop of domain II (C910L), was >10 3 -fold less sensitive to GVIIJ SSG than was wild-type rNa V 1.2. In contrast, although rNa V 1.5 was >10 4 -fold less sensitive to GVIIJ SSG than Na V 1.2, an rNa V 1.5 mutant with a cysteine in the homologous location, rNa V 1.5[L869C], was >10 3 -fold more sensitive than wildtype rNa V 1.5. The susceptibility of rNa V 1.2 to GVIIJ SSG was significantly altered by treating the channels with thiol-oxidizing or disulfide-reducing agents. Furthermore, coexpression of rNa V β2 or rNa V β4, but not that of rNa V β1 or rNa V β3, protected rNa V 1.1 to -1.7 (excluding Na V 1.5) against block by GVIIJ SSG . Thus, GVIIJrelated peptides may serve as probes for both the redox state of extracellular cysteines and for assessing which Na V β-and Na V α-subunits are present in native neurons.oltage-gated sodium channels (VGSCs) are responsible for the upstroke of action potentials in excitable tissues. Each VGSC is composed of a pore-and voltage sensor-bearing α-subunit and one or more auxiliary β-subunits. Mammals have nine α-subunit isoforms (Na V 1.1 to -1.9) and four β-subunit isoforms (Na V β1 to -β4) (1). An Na V 1 has about 2,000-aa residues arranged in four homologous domains, where each domain has six transmembrane spanning segments with an extracellular "pore" loop between segments 5 and 6 (1, 2); furthermore, each Na V 1 has about a dozen extracellular cysteine residues, all located in or near the pore loops. For the most part, not much is known about these cysteines (including whether they are disulfide bonded).Na V β-subunits can affect the function and cellular localization of Na V 1s (1, 3-5). Each Na V β-subunit has some 200-aa residues and consists of a single transmembrane segment with a large extracellular domain and a smaller intracellular domain (1). Na V β2-and Na V β4-subunits, unlike Na V β1-and Na V β3-subunits, are disulfide bonded to α-subunits (1, 6). A given neuron can have multiple isoforms of these subunits whose identities are challenging to appraise pharmacologically (7).Toxins that target VGSCs have been invaluable for probing the structure and function of these channels. Venoms are a rich source of such toxins. For example, in Conus snails, four families of neuroactive peptides have been characterized that target VGSCs:...

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Section: Discussionmentioning

confidence: 99%

A disulfide tether stabilizes the block of sodium channels by the conotoxin μO§-GVIIJ

Gajewiak¹,

Azam²,

Imperial³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…19 The exact pathways leading to the dysregulation of NaV1.7 are poorly understood but likely involve mechanisms related to its surface trafficking and regulation via protein–protein interactions. 18,20–22 Recent studies have identified neuronal CRMP2 as a novel binding partner of NaV1.7. 21,22 Specifically, a selective reduction in NaV1.7 surface expression and current density was observed in rodent and human sensory neurons expressing a mutant CRMP2 lacking the SUMO PTM site (lysine 374) in CRMP2.…”

mentioning

confidence: 99%

Blocking CRMP2 SUMOylation reverses neuropathic pain

et al. 2017

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“…The acceleration of inactivation and the recovery from inactivation of the Na + channel is likely due to increased β3 subunit expression, although steady-state activation and inactivation of this channel were not changed in FoxO1 -/- [31]. β3 regulating Na V 1.5 may have similar mechanisms by which β3 increases both expression of the core-glycosylated form of Na V 1.7 and Na + channel activity [33]. …”

Section: Discussionmentioning

confidence: 99%

Deletion of FoxO1 leads to shortening of QRS by increasing Na+ channel activity through enhanced expression of both cardiac NaV1.5 and β3 subunit

Cai

Wang

Mao

et al. 2014

Journal of Molecular and Cellular Cardiology

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Our in vitro studies revealed that a transcription factor, Forkhead box protein O1 (FoxO1), negatively regulates the expression of NaV1.5, a main α subunit of the cardiac Na+ channel, by altering the promoter activity of SCN5a in HL-1 cardiomyocytes. The in vivo role of FoxO1 in the regulation of cardiac NaV1.5 expression remains unknown. The present study aimed to define the role of FoxO1 in the regulation of NaV1.5 expression and cardiac Na+ channel activity in mouse ventricular cardiomyocytes and assess the cardiac electrophysiological phenotype of mice with cardiac FoxO1 deletion. Tamoxifen-induced and cardiac-specific FoxO1 deletion was confirmed by polymerase chain reaction (PCR). Cardiac FoxO1 deletion failed to result in either cardiac functional changes or hypertrophy as assessed by echocardiography and individual ventricular cell capacitances, respectively. Western blotting showed that FoxO1 was significantly decreased while NaV1.5 protein level was significantly increased in mouse hearts with FoxO1 deletion. Reverse transcription-PCR (RT-PCR) revealed that FoxO1 deletion led to an increase in NaV1.5 and Na+ channel subunit β3 mRNA, but not β1, 2, 4, or connexin 43. Whole patch-clamp recordings demonstrated that cardiac Na+ currents were significantly augmented by FoxO1 deletion without affecting the steady-state activation and inactivation, leading to accelerated depolarization of action potentials in mouse ventricular cardiomyocytes. Electrocardiogram recordings showed that the QRS complex was significantly shortened and P wave amplitude was significantly increased in conscious and unrestrained mice with cardiac FoxO1 deletion. NaV1.5 expression was decreased in the peri-infarct (border-zone) of mice with myocardial infarction and FoxO1 accumulated in the cardiomyocyte nuclei of chronic ischemic human hearts. Our findings indicate that FoxO1 plays an important role in the regulation of NaV1.5 and β3 subunit expression as well as Na+ channel activity in the heart and that FoxO1 is involved in the modulation of NaV1.5 expression in ischemic heart disease.

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β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Nav1.7 in HEK293 cells

Cited by 61 publications

References 47 publications

A disulfide tether stabilizes the block of sodium channels by the conotoxin μO§-GVIIJ

A disulfide tether stabilizes the block of sodium channels by the conotoxin μO§-GVIIJ

Blocking CRMP2 SUMOylation reverses neuropathic pain

Deletion of FoxO1 leads to shortening of QRS by increasing Na+ channel activity through enhanced expression of both cardiac NaV1.5 and β3 subunit

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