title: Intracellular gate opening in Shaker K+ channels defined by high-affinity metal bridges.
abstract: Voltage-gated potassium channels such as Shaker help to control electrical signalling in neurons by regulating the passage of K+ across cell membranes. Ion flow is controlled by a voltage-dependent gate at the intracellular side of the pore, formed by the crossing of four alpha-helices--the inner-pore helices. The prevailing model of gating is based on a comparison of the crystal structures of two bacterial channels--KcsA in a closed state and MthK in an open state--and proposes a hinge motion at a conserved glycine that splays the inner-pore helices wide open. We show here that two types of intersubunit metal bridge, involving cysteines placed near the bundle crossing, can occur simultaneously in the open state. These bridges provide constraints on the open Shaker channel structure, and on the degree of movement upon opening. We conclude that, unlike predictions from the structure of MthK, the inner-pore helices of Shaker probably maintain the KcsA-like bundle-crossing motif in the open state, with a bend in this region at the conserved proline motif (Pro-X-Pro) not found in the bacterial channels. A narrower opening of the bundle crossing in Shaker K+ channels may help to explain why Shaker has an approximately tenfold lower conductance than its bacterial relatives.
authors.affiliation: Department of Neurobiology, Harvard Medical School, 220 Longwood Avenue, Boston, Massachusetts 02115, USA.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Webster
authors.forename: Donato
authors.initials: D
authors.lastname: Del Camino
authors.forename: John P
authors.initials: JP
authors.lastname: Dekker
authors.forename: Gary
authors.initials: G
authors.lastname: Yellen
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002104:Cadmium / Q000378:metabolism*; D002460:Cell Line; D004553:Electric Conductivity; D006801:Humans; D015640:Ion Channel Gating; D017136:Ion Transport; D008024:Ligands; D008958:Models, Molecular; D009154:Mutation; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020836:Protein Structure, Quaternary; D051637:Shaker Superfamily of Potassium Channels
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D015221:Potassium Channels; D051637:Shaker Superfamily of Potassium Channels; C107790:prokaryotic potassium channel; D002104:Cadmium; D011188:Potassium