title: An artificial tetramerization domain restores efficient assembly of functional Shaker channels lacking T1.
abstract: One feature shared by all Shaker-type voltage-gated K(+) channels is a highly conserved domain (T1) located in the cytoplasmic N terminus. The T1 domain is a key determinant of which subtypes can form heteromultimeric channels, suggesting that T1 functions during channel assembly. To better define the role of T1 during channel assembly and separate this function from potential contributions to channel permeation and gating, we replaced the T1 domain (residues 96-183) of ShakerB with a coiled-coil sequence (GCN4-LI) that forms parallel tetramers. Deleting T1 dramatically, but not completely, abolished channel formation under most expression conditions. Channels lacking T1 are functional and K(+)-selective, although they activate at more hyperpolarized membrane potentials and inactivate less completely. Insertion of the artificial tetramerization domain (GCN4-LI) restored efficient channel formation, suggesting that tetramerization of the cytoplasmic T1 domain promotes transmembrane channel assembly by increasing the effective local subunit concentration for T1 compatible subunits. We propose that T1 tetramerization promotes subfamily-specific assembly through kinetic partitioning of the assembly process, but is not required for subsequent steps in channel assembly and folding.
authors.affiliation: Howard Hughes Medical Institute, Departments of Physiology and Biochemistry, Program in Neuroscience, University of California, San Francisco, CA 94143-0725, USA.
authors.forename: N
authors.initials: N
authors.lastname: Zerangue
authors.forename: Y N
authors.initials: YN
authors.lastname: Jan
authors.forename: L Y
authors.initials: LY
authors.lastname: Jan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001704:Biopolymers; D008564:Membrane Potentials / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D051637:Shaker Superfamily of Potassium Channels; D014981:Xenopus
chemical_list: D001704:Biopolymers; D015221:Potassium Channels; D051637:Shaker Superfamily of Potassium Channels