title: Intracellular and extracellular amino acids that influence C-type inactivation and its modulation in a voltage-dependent potassium channel.
abstract: The rate of C-type inactivation of the cloned voltage-gated potassium channel, Kv1.3, measured in membrane patches from Xenopus oocytes, increases when the patch is detached from the cell; the structural basis for this on-cell/off-cell change was examined. First, four serine and threonine residues, that are putative sites for phosphorylation by protein kinases A and C, were mutated to alanines. Mutating any one of these residues, or two or three of them simultaneously, does not eliminate the change in C-type inactivation. However, the basal rate of C-type inactivation in the cell-attached patch is markedly slower in the triple phosphorylation site mutant. Second, a homologous potassium channel, Kv 1.6, does not exhibit the on-cell/off-cell change. When an extracellular histidine at position 401 of Kv1.3 is replaced with tyrosine, the residue at the equivalent position (430) in Kv1.6, the resulting Kv1.3 H401Y mutant channel does not undergo the on-cell/off-cell change. The results indicate that several potentially phosphorylatable intracellular amino acids influence the basal rate of C-type inactivation, but are not essential for the on-cell/off-cell change in inactivation kinetics. In contrast, an extracellular amino acid is critical for this on-cell/off-cell change.
authors.affiliation: Biochemistry Department, Brandeis University, Waltham MA 02254, USA.
authors.forename: J
authors.initials: J
authors.lastname: Kupper
authors.forename: M R
authors.initials: MR
authors.lastname: Bowlby
authors.forename: S
authors.initials: S
authors.lastname: Marom
authors.forename: I B
authors.initials: IB
authors.lastname: Levitan
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D005260:Female; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D008564:Membrane Potentials / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D010766:Phosphorylation; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D013329:Structure-Activity Relationship; D014981:Xenopus
chemical_list: D000596:Amino Acids; D015221:Potassium Channels