title: Probing the transition state of the allosteric pathway of the Shaker Kv channel pore by linear free-energy relations.
abstract: Long-range coupling between distant functional elements of proteins may rely on allosteric communication trajectories lying along the protein structure, as described in the case of the Shaker voltage-activated potassium (Kv) channel model allosteric system. Communication between the distant Kv channel activation and slow inactivation pore gates was suggested to be mediated by a network of local pairwise and higher-order interactions among the functionally unique residues that constitute the allosteric trajectory. The mechanism by which conformational changes propagate along the Kv channel allosteric trajectory to achieve pore opening, however, remains unclear. Such conformational changes may propagate in either a concerted or a sequential manner during the reaction coordinate of channel opening. Residue-level structural information on the transition state of channel gating is required to discriminate between these possibilities. Here, we combine patch-clamp electrophysiology recordings of Kv channel gating and analysis using linear free-energy relations, focusing on a select set of residues spanning the allosteric trajectory of the Kv channel pore. We show that all allosteric trajectory residues tested exhibit an open-like conformation in the transition state of channel opening, implying that coupling interactions occur along the trajectory break in a concerted manner upon moving from the closed to the open state. Energetic coupling between the Kv channel gates thus occurs in a concerted fashion in both the spatial and the temporal dimensions, strengthening the notion that such trajectories correspond to pathways of mechanical deformation along which conformational changes propagate.
authors.affiliation: Department of Life Sciences and the Zlotowski Center for Neurosciences, Ben-Gurion University of the Negev, PO Box 653, Beer Sheva 84105, Israel.
authors.forename: Reshef
authors.initials: R
authors.lastname: Azaria
authors.forename: Orr
authors.initials: O
authors.lastname: Irit
authors.forename: Yuval
authors.initials: Y
authors.lastname: Ben-Abu
authors.forename: Ofer
authors.initials: O
authors.lastname: Yifrach
mesh_terms: D000494:Allosteric Regulation; D015640:Ion Channel Gating; D008954:Models, Biological; D008958:Models, Molecular; D018408:Patch-Clamp Techniques; D051637:Shaker Superfamily of Potassium Channels / Q000737:chemistry* / Q000378:metabolism*; D013997:Time Factors
chemical_list: D051637:Shaker Superfamily of Potassium Channels