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Crystal structure and molecular imaging of the Nav channel β3 subunit indicates a trimeric assembly.

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cam.issuedOnline2014-02-24
dc.contributor.authorNamadurai, Sivakumar
dc.contributor.authorBalasuriya, Dilshan
dc.contributor.authorRajappa, Rajit
dc.contributor.authorWiemhöfer, Martin
dc.contributor.authorStott, Katherine
dc.contributor.authorKlingauf, Jurgen
dc.contributor.authorEdwardson, J Michael
dc.contributor.authorChirgadze, Dimitri Y
dc.contributor.authorJackson, Antony P
dc.contributor.orcidStott, Katherine [0000-0002-4014-1188]
dc.contributor.orcidChirgadze, Dima [0000-0001-9942-0993]
dc.contributor.orcidJackson, Antony [0000-0002-2895-7387]
dc.date.accessioned2018-09-20T12:04:16Z
dc.date.available2018-09-20T12:04:16Z
dc.date.issued2014-04-11
dc.description.abstractThe vertebrate sodium (Nav) channel is composed of an ion-conducting α subunit and associated β subunits. Here, we report the crystal structure of the human β3 subunit immunoglobulin (Ig) domain, a functionally important component of Nav channels in neurons and cardiomyocytes. Surprisingly, we found that the β3 subunit Ig domain assembles as a trimer in the crystal asymmetric unit. Analytical ultracentrifugation confirmed the presence of Ig domain monomers, dimers, and trimers in free solution, and atomic force microscopy imaging also detected full-length β3 subunit monomers, dimers, and trimers. Mutation of a cysteine residue critical for maintaining the trimer interface destabilized both dimers and trimers. Using fluorescence photoactivated localization microscopy, we detected full-length β3 subunit trimers on the plasma membrane of transfected HEK293 cells. We further show that β3 subunits can bind to more than one site on the Nav 1.5 α subunit and induce the formation of α subunit oligomers, including trimers. Our results suggest a new and unexpected role for the β3 subunits in Nav channel cross-linking and provide new structural insights into some pathological Nav channel mutations.
dc.format.mediumPrint-Electronic
dc.identifier.doi10.17863/CAM.27837
dc.identifier.eissn1083-351X
dc.identifier.issn0021-9258
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/280466
dc.languageeng
dc.language.isoeng
dc.publisherElsevier BV
dc.publisher.urlhttp://dx.doi.org/10.1074/jbc.m113.527994
dc.subjectAnalytical Ultracentrifugation
dc.subjectAtomic Force Microscopy
dc.subjectIn Vivo Imaging
dc.subjectPhotoactivated Localization Microscopy
dc.subjectProtein Trimerization
dc.subjectSodium Channels
dc.subjectX-ray Crystallography
dc.subjectAmino Acid Sequence
dc.subjectBinding Sites
dc.subjectCloning, Molecular
dc.subjectCrystallization
dc.subjectCrystallography, X-Ray
dc.subjectDimerization
dc.subjectHEK293 Cells
dc.subjectHumans
dc.subjectImmunoglobulins
dc.subjectMicroscopy, Atomic Force
dc.subjectMolecular Sequence Data
dc.subjectNAV1.5 Voltage-Gated Sodium Channel
dc.subjectProtein Conformation
dc.subjectUltracentrifugation
dc.subjectVoltage-Gated Sodium Channel beta-3 Subunit
dc.titleCrystal structure and molecular imaging of the Nav channel β3 subunit indicates a trimeric assembly.
dc.typeArticle
prism.endingPage10811
prism.issueIdentifier15
prism.publicationDate2014
prism.publicationNameJ Biol Chem
prism.startingPage10797
prism.volume289
pubs.funder-project-idWellcome Trust (089125/Z/09/Z)
rioxxterms.licenseref.startdate2014-04
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionAM
rioxxterms.versionofrecord10.1074/jbc.M113.527994

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