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The Journal of Neuroscience, February 15, 2001, 21(4):1160-1168
Casein Kinase 2 Determines the Voltage Dependence of the Kv3.1
Channel in Auditory Neurons and Transfected Cells
Carolyn M.
Macica and
Leonard K.
Kaczmarek
Department of Pharmacology, Yale University School of Medicine, New
Haven, Connecticut 06520-8066
The Kv3.1 potassium channel can be distinguished from most other
delayed rectifier channels by its very high threshold of activation and
lack of use-dependent inactivation. This allows neurons that express
this channel to fire at very high frequencies. We have now found that
this feature of the Kv3.1 channel is strongly influenced by its
constitutive phosphorylation by the enzyme casein kinase II. Using
stably transfected Chinese hamster ovary cells expressing Kv3.1,
we show that Kv3.1 is highly phosphorylated under basal conditions.
Whole-cell patch clamp recordings were used to characterize the
electrophysiological consequence of dephosphorylation using alkaline
phosphatase. This enzyme produced an increase in whole-cell conductance
and shifted the voltage dependence of activation to more negative
potentials by >20 mV. In addition, a similar shift in the voltage
dependence of inactivation was observed. These findings were also
confirmed in native Kv3.1 channels expressed in medial nucleus of the
trapezoid body (MNTB) neurons. Furthermore, inhibitors of casein kinase
2 mimicked the effect of phosphatase treatment on voltage-dependent
activation and inactivation, whereas inhibitors of protein kinase
C failed to alter these parameters. The combination of
biochemical and electrophysiological evidence suggests that the
biophysical characteristics of Kv3.1 that are important to its role in
MNTB neurons, allowing them to follow high-frequency stimuli with
fidelity, are largely determined by phosphorylation of the channel.
Key words:
Kv3.1; potassium channel; constitutive phosphorylation; casein kinase; MNTB neuron; voltage dependence of activation; voltage
dependence of inactivation
Copyright © 2001 Society for Neuroscience 0270-6474/01/2141160-09$05.00/0
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