Dataset Information

Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel ? subunits.

ABSTRACT: The ?1, ?2, and ?4 subunits of voltage-gated sodium channels reportedly function as cell adhesion molecules. The present crystallographic analysis of the ?4 extracellular domain revealed an antiparallel arrangement of the ?4 molecules in the crystal lattice. The interface between the two antiparallel ?4 molecules is asymmetric, and results in a multimeric assembly. Structure-based mutagenesis and site-directed photo-crosslinking analyses of the ?4-mediated cell-cell adhesion revealed that the interface between the antiparallel ?4 molecules corresponds to that in the trans homophilic interaction for the multimeric assembly of ?4 in cell-cell adhesion. This trans interaction mode is also employed in the ?1-mediated cell-cell adhesion. Moreover, the ?1 gene mutations associated with generalized epilepsy with febrile seizures plus (GEFS+) impaired the ?1-mediated cell-cell adhesion, which should underlie the GEFS+ pathogenesis. Thus, the structural basis for the ?-subunit-mediated cell-cell adhesion has been established.

SUBMITTER: Shimizu H

PROVIDER: S-EPMC4877568 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel β subunits.

Shimizu Hideaki H Miyazaki Haruko H Ohsawa Noboru N Shoji Shisako S Ishizuka-Katsura Yoshiko Y Tosaki Asako A Oyama Fumitaka F Terada Takaho T Sakamoto Kensaku K Shirouzu Mikako M Sekine Shun-Ichi S Nukina Nobuyuki N Yokoyama Shigeyuki S

Scientific reports 20160524

The β1, β2, and β4 subunits of voltage-gated sodium channels reportedly function as cell adhesion molecules. The present crystallographic analysis of the β4 extracellular domain revealed an antiparallel arrangement of the β4 molecules in the crystal lattice. The interface between the two antiparallel β4 molecules is asymmetric, and results in a multimeric assembly. Structure-based mutagenesis and site-directed photo-crosslinking analyses of the β4-mediated cell-cell adhesion revealed that the in ...[more]

PMID: 27216889

Similar Datasets

Project description:Background:Voltage-gated sodium (NaV) channels are heteromeric proteins consisting of a single pore forming ?-subunit associated with one or two auxiliary ?-subunits. These channels are classically known for being responsible of action potential generation and propagation in excitable cells; but lately they have been reported as widely expressed and regulated in several human cancer types. We have previously demonstrated the overexpression of NaV1.6 channel in cervical cancer (CeCa) biopsies and primary cultures, and its contribution to cell migration and invasiveness. Here, we investigated the expression of NaV channels ?-subunits (NaV?s) in the CeCa cell lines HeLa, SiHa and CaSki, and determined their contribution to cell proliferation, migration and invasiveness. Methods:We assessed the expression of NaV?s in CeCa cell lines by performing RT-PCR and western blotting experiments. We also evaluated CeCa cell lines proliferation, migration, and invasion by in vitro assays, both in basal conditions and after inducing changes in NaV?s levels by transfecting specific cDNAs or siRNAs. The potential role of NaV?s in modulating the expression of NaV ?-subunits in the plasma membrane of CeCa cells was examined by the patch-clamp whole-cell technique. Furthermore, we investigated the role of NaV?1 on cell cycle in SiHa cells by flow cytometry. Results:We found that the four NaV?s are expressed in the three CeCa cell lines, even in the absence of functional NaV ?-subunit expression in the plasma membrane. Functional in vitro assays showed differential roles for NaV?1 and NaV?4, the latter as a cell invasiveness repressor and the former as a migration abolisher in CeCa cells. In silico analysis of NaV?4 expression in cervical tissues corroborated the downregulation of this protein expression in CeCa vs normal cervix, supporting the evidence of NaV?4's role as a cell invasiveness repressor. Conclusions:Our results contribute to the recent conception about NaV?s as multifunctional proteins involved in cell processes like ion channel regulation, cell adhesion and motility, and even in metastatic cell behaviors. These non-canonical functions of NaV?s are independent of the presence of functional NaV ?-subunits in the plasma membrane and might represent a new therapeutic target for the treatment of cervical cancer.

Dataset Information

Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel ? subunits.

Publications

Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel β subunits.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets