title: Benchmarking the stability of human detergent-solubilised voltage-gated sodium channels for structural studies using eel as a reference.
abstract: With the ultimate goal of detailed structural analysis of mammalian and particularly human voltage-gated sodium channels (VGSCs), we have investigated the relative stability of human and rat VGSCs and compared them with electric eel VGSC. We found that NaV1.3 from rat was the most stable after detergent solubilisation. The order of stability was rNaV1.3>hNaV1.2>hNaV1.1>hNaV1.6>hNaV1.3>hNaV1.4. However, a comparison with the VGSC from Electrophorus electricus, which is most similar to NaV1.4, shows that the eel VGSC is considerably more stable in detergent than the human VGSCs examined. We conclude that current methods of structural analysis, such as single particle electron cryomicroscopy (cryoEM), may be most usefully targeted to eel VGSC or rNaV1.3, but that structural analysis on the full spectrum of VGSCs, by methods that require greater stability such as crystallisation and X-ray crystallography, will require further stabilisation of the channel.
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom. Electronic address: dslowik@mrc-lmb.cam.ac.uk.
authors.forename: Daria
authors.initials: D
authors.lastname: Slowik
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom. Electronic address: rh15@mrc-lmb.cam.ac.uk.
authors.forename: Richard
authors.initials: R
authors.lastname: Henderson
mesh_terms: D000818:Animals; D019985:Benchmarking / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D003902:Detergents / Q000737:chemistry*; D004524:Eels / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D062550:NAV1.1 Voltage-Gated Sodium Channel / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D062551:NAV1.2 Voltage-Gated Sodium Channel / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D062552:NAV1.3 Voltage-Gated Sodium Channel / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D055550:Protein Stability / Q000187:drug effects; D051381:Rats; D012995:Solubility; D013696:Temperature; D014118:Toxins, Biological / Q000378:metabolism / Q000494:pharmacology; D061566:Voltage-Gated Sodium Channels / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D062550:NAV1.1 Voltage-Gated Sodium Channel; D062551:NAV1.2 Voltage-Gated Sodium Channel; D062552:NAV1.3 Voltage-Gated Sodium Channel; C568242:SCN1A protein, human; C478638:Scn3a protein, rat; D014118:Toxins, Biological; D061566:Voltage-Gated Sodium Channels
keywords: Electrophorus electricus; Human voltage-gated sodium channel; Membrane protein stability; Radioligand-binding assay; Rat VGSC; Structural biology
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