title: A carboxy-terminal alpha-helical segment in the rat skeletal muscle voltage-dependent Na+ channel is responsible for its interaction with the amino-terminus.
abstract: Cytoplasmic segments of the adult rat skeletal muscle sodium channel alpha-subunit (rSkM1) comprise a major portion (approximately 40%) of the total protein and are involved in channel functions both general, such as inactivation, and isoform-specific, for example, protein kinase A modulation. Far ultraviolet circular dichroism measurements of synthetic peptides and overexpressed fusion proteins containing individual channel cytoplasmic segments suggest that cytoplasmic domains of rSkM1 contain ordered secondary structures even in the absence of adjoining transmembrane segments. Intrinsic fluorescence experiments with a nested set of carboxy-terminal deletion proteins confirm a specific interaction between the channel's amino- and carboxy-termini and identify residues 1716-1737 in the carboxy-terminus as the region that binds to the amino-terminus. Circular dichroism measurements suggest that this same region is organized as an alpha-helix and that electrostatic forces may contribute to this association. The interaction of the amino- and carboxy-termini is not accompanied by secondary structure changes detectable by circular dichroism spectroscopy, but a decrease in intrinsic fluorescence indicates that this association is accompanied by a change in the environment of Trp1617.
authors.affiliation: Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia 19104-6059, USA.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: S
authors.initials: S
authors.lastname: Kolibal
authors.forename: J M
authors.initials: JM
authors.lastname: Vanderkooi
authors.forename: S A
authors.initials: SA
authors.lastname: Cohen
authors.forename: R G
authors.initials: RG
authors.lastname: Kallen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics; D002942:Circular Dichroism; D017931:DNA Primers / Q000235:genetics; D066298:In Vitro Techniques; D058619:Maltose-Binding Proteins; D018482:Muscle, Skeletal / Q000737:chemistry*; D009842:Oligopeptides; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D017384:Sequence Deletion; D015222:Sodium Channels / Q000737:chemistry* / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D058619:Maltose-Binding Proteins; D009842:Oligopeptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D015222:Sodium Channels; C085706:FLAG peptide