Changes in activation gating of IsK potassium currents brought about by mutations in the transmembrane sequence.
Expression of the rat kidney IsK protein in Xenopus oocytes produces slowly-activating potassium channel currents. We have investigated the relationship between structure and function of the single putative membrane-spanning domain using site-directed mutagenesis. Six mutants were constructed in which consecutive individual amino acids (53 to 58) of the transmembrane region were substituted by cysteine. Expression of four of these mutants in Xenopus oocytes resulted in currents which were similar to wild-type. However, for one mutant (position 55) activation curves were shifted in a hyperpolarising direction and for another mutant (position 58) activation curve were shifted in a depolarising direction. This suggests that the hydrophobic phenylalanine residues at positions 55 and 58 may play a critical role in IsK activation gating. This spacing of functional amino acids at every third residue may indicate an alpha-helical conformation for the membrane-spanning domain of IsK. Furthermore, these results also indicate that one face of the helix may represent a region of subunit association.
Wilson
G G
GG
Department of Pharmacology, University of Leeds, UK.
Sivaprasadarao
A
A
Findlay
J B
JB
Wray
D
D
D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000502:physiology*; D015640:Ion Channel Gating / Q000502:physiology*; D007668:Kidney; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000502:physiology; D009865:Oocytes / Q000378:metabolism; D010649:Phenylalanine / Q000502:physiology; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D024642:Potassium Channels, Voltage-Gated; D017433:Protein Structure, Secondary; D051381:Rats; D014981:Xenopus
D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; C059983:potassium channel protein I(sk); D010649:Phenylalanine