jShaw1, a low-threshold, fast-activating Kv3 from the hydrozoan jellyfish Polyorchis penicillatus

Abstract

SUMMARY Voltage-gated potassium (Kv) channels work in concert with other ion channels to determine the frequency and duration of action potentials in excitable cells. Little is known about Kv3 channels from invertebrates, but those that have been characterized generally display slow kinetics. Here, we report the cloning and characterization of jShaw1, the first Kv3 isolated from a cnidarian, the jellyfish Polyorchis penicillatus, in comparison with mouse Kv3.1 and Kv3.2. Using a two-electrode voltage clamp on Xenopus laevis oocytes expressing the channels, we compared steady-state and kinetic properties of macroscopic currents. jShaw1 is fast activating, and opens at potentials approximately 40 mV more hyperpolarized than the mouse Kv3 channels. There is an inverse relationship between the number of positive charges on the voltage sensor and the half-activation voltage of the channel, contrary to what would be expected with the simplest model of voltage sensitivity. jShaw1 has kinetic characteristics that are substantially different from the mammalian Kv3 channels, including a much lower sensitivity of early activation rates to incremental voltage changes, and a much faster voltage-dependent transition in the last stages of opening. jShaw1 opening kinetics were affected little by pre-depolarization voltage, in contrast to both mouse channels. Similar to the mouse channels, jShaw1 was half-blocked by 0.7 mmol l–1 tetraethyl ammonium and 5 mmol l–1 4-aminopyridine. Comparison of sequence and functional properties of jShaw1 with the mouse and other reported Kv3 channels helps to illuminate the general relationship between amino acid sequence and electrophysiological activity in this channel family.