Cloning of a putative voltage-gated sodium channel from the turbellarian flatworm Bdelloura candida

Parasitology ◽  
1997 ◽  
Vol 115 (3) ◽  
pp. 289-296 ◽  
Author(s):  
M. C. JEZIORSKI ◽  
R. M. GREENBERG ◽  
P. A. V. ANDERSON
Keyword(s):  
Sodium Channels ◽  
Northern Blot Analysis ◽  
Amino Acid Identity ◽  
Structure Characteristic ◽  
Sodium Currents ◽  
Voltage Gated ◽  
Transmembrane Segments ◽  
Voltage Gated Sodium Channels ◽  
Multidomain Structure ◽  
Insight Into

The neuromuscular sodium currents of early invertebrates such as platyhelminths display distinctive kinetic and pharmacological properties. We have cloned a cDNA from the horseshoe crab flatworm Bdelloura candida that encodes a protein homologous to the primary subunit of voltage-gated sodium channels. The B. candida protein, named BdNa1, exhibits amino acid identity of 40–47% to sodium channels of vertebrates and higher invertebrates. BdNa1 has the multidomain structure characteristic of sodium channels, and is most highly conserved in the hydrophobic transmembrane segments and the regions that form the pore of the channel. Northern blot analysis confirms the presence of a 5·4 kb BdNa1 transcript in B. candida tissue. The information provided by analysis of the BdNa1 sequence offers insight into the physiology of platyhelminth sodium currents.

scholarly journals Neurosteroids Allopregnanolone Sulfate and Pregnanolone Sulfate Have Diverse Effect on the α Subunit of the Neuronal Voltage-gated Sodium Channels Nav1.2, Nav1.6, Nav1.7, and Nav1.8 Expressed in Xenopus Oocytes

2014 ◽  
Vol 121 (3) ◽  
pp. 620-631 ◽  
Author(s):  
Takafumi Horishita ◽  
Nobuyuki Yanagihara ◽  
Susumu Ueno ◽  
Yuka Sudo ◽  
Yasuhito Uezono ◽  
...  
Keyword(s):  
Sodium Channels ◽  
Xenopus Oocytes ◽  
Sodium Current ◽  
Dependent Manner ◽  
Sodium Currents ◽  
Concentration Dependent Manner ◽  
Α Subunit ◽  
Voltage Gated ◽  
Voltage Gated Sodium Channels ◽  
Α Subunits

Abstract Background: The neurosteroids allopregnanolone and pregnanolone are potent positive modulators of γ-aminobutyric acid type A receptors. Antinociceptive effects of allopregnanolone have attracted much attention because recent reports have indicated the potential of allopregnanolone as a therapeutic agent for refractory pain. However, the analgesic mechanisms of allopregnanolone are still unclear. Voltage-gated sodium channels (Nav) are thought to play important roles in inflammatory and neuropathic pain, but there have been few investigations on the effects of allopregnanolone on sodium channels. Methods: Using voltage-clamp techniques, the effects of allopregnanolone sulfate (APAS) and pregnanolone sulfate (PAS) on sodium current were examined in Xenopus oocytes expressing Nav1.2, Nav1.6, Nav1.7, and Nav1.8 α subunits. Results: APAS suppressed sodium currents of Nav1.2, Nav1.6, and Nav1.7 at a holding potential causing half-maximal current in a concentration-dependent manner, whereas it markedly enhanced sodium current of Nav1.8 at a holding potential causing maximal current. Half-maximal inhibitory concentration values for Nav1.2, Nav1.6, and Nav1.7 were 12 ± 4 (n = 6), 41 ± 2 (n = 7), and 131 ± 15 (n = 5) μmol/l (mean ± SEM), respectively. The effects of PAS were lower than those of APAS. From gating analysis, two compounds increased inactivation of all α subunits, while they showed different actions on activation of each α subunit. Moreover, two compounds showed a use-dependent block on Nav1.2, Nav1.6, and Nav1.7. Conclusion: APAS and PAS have diverse effects on sodium currents in oocytes expressing four α subunits. APAS inhibited the sodium currents of Nav1.2 most strongly.

scholarly journals Molecular Determinants of Brevetoxin Binding to Voltage-Gated Sodium Channels

Toxins ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 513 ◽  
Author(s):  
Keiichi Konoki ◽  
Daniel G. Baden ◽  
Todd Scheuer ◽  
William A. Catterall
Keyword(s):  
Sodium Channels ◽  
Red Tides ◽  
Shellfish Poisoning ◽  
Alanine Scanning Mutagenesis ◽  
Α Subunit ◽  
Voltage Gated ◽  
Transmembrane Segments ◽  
Voltage Gated Sodium Channels ◽  
Fold Reduction ◽  
Molecular Determinants

Brevetoxins are produced by dinoflagellates such as Karenia brevis in warm-water red tides and cause neurotoxic shellfish poisoning. They bind to voltage-gated sodium channels at neurotoxin receptor 5, making the channels more active by shifting the voltage-dependence of activation to more negative potentials and by slowing the inactivation process. Previous work using photoaffinity labeling identified binding to the IS6 and IVS5 transmembrane segments of the channel α subunit. We used alanine-scanning mutagenesis to identify molecular determinants for brevetoxin binding in these regions as well as adjacent regions IVS5-SS1 and IVS6. Most of the mutant channels containing single alanine substitutions expressed functional protein in tsA-201 cells and bound to the radioligand [42-3H]-PbTx3. Binding affinity for the great majority of mutant channels was indistinguishable from wild type. However, transmembrane segments IS6, IVS5 and IVS6 each contained 2 to 4 amino acid positions where alanine substitution resulted in a 2–3-fold reduction in brevetoxin affinity, and additional mutations caused a similar increase in brevetoxin affinity. These findings are consistent with a model in which brevetoxin binds to a protein cleft comprising transmembrane segments IS6, IVS5 and IVS6 and makes multiple distributed interactions with these α helices. Determination of brevetoxin affinity for Nav1.2, Nav1.4 and Nav1.5 channels showed that Nav1.5 channels had a characteristic 5-fold reduction in affinity for brevetoxin relative to the other channel isoforms, suggesting the interaction with sodium channels is specific despite the distributed binding determinants.

scholarly journals Voltage-gated sodium channels (version 2019.4) in the IUPHAR/BPS Guide to Pharmacology Database

2019 ◽  
Vol 2019 (4) ◽  
Author(s):  
William A. Catterall ◽  
Alan L. Goldin ◽  
Stephen G. Waxman
Keyword(s):  
Sodium Channels ◽  
Ion Selectivity ◽  
Structural Features ◽  
Fatty Acyl ◽  
Transmembrane Segment ◽  
Α Subunit ◽  
Voltage Gated ◽  
Transmembrane Segments ◽  
Voltage Gated Sodium Channels ◽  
Acyl Chains

Sodium channels are voltage-gated sodium-selective ion channels present in the membrane of most excitable cells. Sodium channels comprise of one pore-forming α subunit, which may be associated with either one or two β subunits [176]. α-Subunits consist of four homologous domains (I–IV), each containing six transmembrane segments (S1–S6) and a pore-forming loop. The positively charged fourth transmembrane segment (S4) acts as a voltage sensor and is involved in channel gating. The crystal structure of the bacterial NavAb channel has revealed a number of novel structural features compared to earlier potassium channel structures including a short selectivity filter with ion selectivity determined by interactions with glutamate side chains [268]. Interestingly, the pore region is penetrated by fatty acyl chains that extend into the central cavity which may allow the entry of small, hydrophobic pore-blocking drugs [268]. Auxiliary β1, β2, β3 and β4 subunits consist of a large extracellular N-terminal domain, a single transmembrane segment and a shorter cytoplasmic domain.The nomenclature for sodium channels was proposed by Goldin et al., (2000) [143] and approved by the NC-IUPHAR Subcommittee on sodium channels (Catterall et al., 2005, [51]).

scholarly journals Voltage-gated sodium channels (NaV) in GtoPdb v.2021.3

2021 ◽  
Vol 2021 (3) ◽  
Author(s):  
William A. Catterall ◽  
Alan L. Goldin ◽  
Stephen G. Waxman
Keyword(s):  
Sodium Channels ◽  
Ion Selectivity ◽  
Structural Features ◽  
Fatty Acyl ◽  
Transmembrane Segment ◽  
Α Subunit ◽  
Voltage Gated ◽  
Transmembrane Segments ◽  
Voltage Gated Sodium Channels ◽  
Acyl Chains

Sodium channels are voltage-gated sodium-selective ion channels present in the membrane of most excitable cells. Sodium channels comprise of one pore-forming α subunit, which may be associated with either one or two β subunits [177]. α-Subunits consist of four homologous domains (I-IV), each containing six transmembrane segments (S1-S6) and a pore-forming loop. The positively charged fourth transmembrane segment (S4) acts as a voltage sensor and is involved in channel gating. The crystal structure of the bacterial NavAb channel has revealed a number of novel structural features compared to earlier potassium channel structures including a short selectivity filter with ion selectivity determined by interactions with glutamate side chains [274]. Interestingly, the pore region is penetrated by fatty acyl chains that extend into the central cavity which may allow the entry of small, hydrophobic pore-blocking drugs [274]. Auxiliary β1, β2, β3 and β4 subunits consist of a large extracellular N-terminal domain, a single transmembrane segment and a shorter cytoplasmic domain.The nomenclature for sodium channels was proposed by Goldin et al., (2000) [144] and approved by the NC-IUPHAR Subcommittee on sodium channels (Catterall et al., 2005, [52]).

scholarly journals α- And β-subunit composition of voltage-gated sodium channels investigated with μ-conotoxins and the recently discovered μO§-conotoxin GVIIJ

2015 ◽  
Vol 113 (7) ◽  
pp. 2289-2301 ◽  
Author(s):  
Michael J. Wilson ◽  
Min-Min Zhang ◽  
Joanna Gajewiak ◽  
Layla Azam ◽  
Jean E. Rivier ◽  
...  
Keyword(s):  
Sciatic Nerve ◽  
Sodium Channels ◽  
Action Potentials ◽  
Xenopus Laevis Oocytes ◽  
Sodium Currents ◽  
Voltage Gated ◽  
Voltage Gated Sodium Channels ◽  
Compound Action Potentials ◽  
Sciatic Nerves ◽  
Synthetic Derivatives

We investigated the identities of the isoforms of the α (NaV1)- and β (NaVβ)-subunits of voltage-gated sodium channels, including those responsible for action potentials in rodent sciatic nerves. To examine α-subunits, we used seven μ-conotoxins, which target site 1 of the channel. With the use of exogenously expressed channels, we show that two of the μ-conotoxins, μ-BuIIIB and μ-SxIIIA, are 50-fold more potent in blocking NaV1.6 from mouse than that from rat. Furthermore, we observed that μ-BuIIIB and μ-SxIIIA are potent blockers of large, myelinated A-fiber compound action potentials (A-CAPs) [but not small, unmyelinated C-fiber CAPs (C-CAPs)] in the sciatic nerve of the mouse (unlike A-CAPs of the rat, previously shown to be insensitive to these toxins). To investigate β-subunits, we used two synthetic derivatives of the recently discovered μO§-conotoxin GVIIJ that define site 8 of the channel, as previously characterized with cloned rat NaV1- and NaVβ-subunits expressed in Xenopus laevis oocytes, where it was shown that μO§-GVIIJ is a potent inhibitor of several NaV1-isoforms and that coexpression of NaVβ2 or -β4 (but not NaVβ1 or -β3) totally protects against block by μO§-GVIIJ. We report here the effects of μO§-GVIIJ on 1) sodium currents of mouse NaV1.6 coexpressed with various combinations of NaVβ-subunits in oocytes; 2) A- and C-CAPs of mouse and rat sciatic nerves; and 3) sodium currents of small and large neurons dissociated from rat dorsal root ganglia. Our overall results lead us to conclude that action potentials in A-fibers of the rodent sciatic nerve are mediated primarily by NaV1.6 associated with NaVβ2 or NaVβ4.

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