title: Mutations in the S6 gate isolate a late step in the activation pathway and reduce 4-AP sensitivity in shaker K(v) channel.
abstract: Kv channels detect changes in the membrane potential via their voltage-sensing domains (VSDs) that control the status of the S6 bundle crossing (BC) gate. The movement of the VSDs results in a transfer of the S4 gating charges across the cell membrane but only the last 10-20% of the total gating charge movement is associated with BC gate opening, which involves cooperative transition(s) in the subunits. Substituting the proline residue P475 in the S6 of the Shaker channel by a glycine or alanine causes a considerable shift in the voltage-dependence of the cooperative transition(s) of BC gate opening, effectively isolating the late gating charge component from the other gating charge that originates from earlier VSD movements. Interestingly, both mutations also abolished Shaker's sensitivity to 4-aminopyridine, which is a pharmacological tool to isolate the late gating charge component. The alanine substitution (that would promote a α-helical configuration compared to proline) resulted in the largest separation of both gating charge components; therefore, BC gate flexibility appears to be important for enabling the late cooperative step of channel opening.
authors.affiliation: Laboratory for Molecular Biophysics, Physiology and Pharmacology, University of Antwerp, Antwerp, Belgium.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Martinez-Morales
authors.affiliation: Laboratory for Molecular Biophysics, Physiology and Pharmacology, University of Antwerp, Antwerp, Belgium.
authors.forename: Dirk J
authors.initials: DJ
authors.lastname: Snyders
authors.affiliation: Laboratory for Molecular Biophysics, Physiology and Pharmacology, University of Antwerp, Antwerp, Belgium. Electronic address: alain.labro@uantwerpen.be.
authors.forename: Alain J
authors.initials: AJ
authors.lastname: Labro
mesh_terms: D015761:4-Aminopyridine / Q000494:pharmacology*; D000595:Amino Acid Sequence; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating; D008969:Molecular Sequence Data; D009154:Mutation; D026902:Potassium Channel Blockers / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D051637:Shaker Superfamily of Potassium Channels / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D026902:Potassium Channel Blockers; D051637:Shaker Superfamily of Potassium Channels; D015761:4-Aminopyridine