Episodic ataxia type 1 mutation F184C alters Zn2+-induced modulation of the human K+ channel Kv1.4-Kv1.1/Kvbeta1.1.
Episodic ataxia type 1 (EA1) is a Shaker-like channelopathy characterized by continuous myokymia and attacks of imbalance with jerking movements of the head, arms, and legs. Although altered expression and gating properties of Kv1.1 channels underlie EA1, several disease-causing mechanisms remain poorly understood. It is likely that Kv1.1, Kv1.4, and Kvbeta1.1 subunits form heteromeric channels at hippocampal mossy fiber boutons from which Zn(2+) ions are released into the synaptic cleft in a Ca(2+)-dependent fashion. The sensitivity of this macromolecular channel complex to Zn(2+) is unknown. Here, we show that this heteromeric channel possesses a high-affinity (<10 muM) and a low-affinity (<0.5 mM) site for Zn(2+), which are likely to regulate channel availability at distinct presynaptic membranes. Furthermore, the EA1 mutation F184C, located within the S1 segment of the Kv1.1 subunit, markedly decreased the equilibrium dissociation constants for Zn(2+) binding to the high- and low-affinity sites. The functional characterization of the Zn(2+) effects on heteromeric channels harboring the F184C mutation also showed that this ion significantly 1) slowed the activation rate of the channel, 2) increased the time to reach peak current amplitude, 3) decreased the rate and amount of current undergoing N-type inactivation, and 4) slowed the repriming of the channel compared with wild-type channels. These results demonstrate that the EA1 mutation F184C will not only sensitize the homomeric Kv1.1 channel to extracellular Zn(2+), but it will also endow heteromeric channels with a higher sensitivity to this metal ion. During the vesicular release of Zn(2+), its effects will be in addition to the intrinsic gating defects caused by the mutation, which is likely to exacerbate the symptoms by impairing the integration and transmission of signals within specific brain areas.
Imbrici
Paola
P
Section of Human Physiology, Dept. of Internal Medicine, Univ. of Perugia School of Medicine, Via del Giochetto, I-06126 Perugia, Italy.
D'Adamo
Maria Cristina
MC
Cusimano
Antonella
A
Pessia
Mauro
M
D000818:Animals; D002118:Calcium / Q000502:physiology; D002413:Cations, Divalent; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D051662:Kv1.1 Potassium Channel / Q000235:genetics / Q000502:physiology*; D051665:Kv1.4 Potassium Channel / Q000235:genetics / Q000502:physiology*; D009154:Mutation; D009865:Oocytes / Q000502:physiology; D017729:Presynaptic Terminals / Q000502:physiology; D013132:Spinocerebellar Degenerations / Q000235:genetics; D014982:Xenopus laevis; D015032:Zinc / Q000494:pharmacology*
D002413:Cations, Divalent; D051665:Kv1.4 Potassium Channel; D051662:Kv1.1 Potassium Channel; D015032:Zinc; D002118:Calcium