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Cyclic AMP-dependent phosphorylation modifies the gating properties of L-type Ca2+ channels in bovine adrenal chromaffin cells

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Abstract

We investigated the effects of cAMP-dependent phosphorylation on the voltage- and time-dependent gating properties of Ca2+ channel currents recorded from bovine adrenal chromaffin cells under whole-cell voltage clamp. Extracellular perfusion with the membrane-permeant activator of cAMP-dependent protein kinase, 8-bromo(8-Br)-cAMP (1 mM), caused a 49%, 29%, and 21% increase in Ca2+ current (I Ca) amplitudes evoked by voltage steps to 0, +10, and +20 mV respectively (mean values from eight cells, p≤0.05). Analysis of voltage-dependent steady-state activation (m ∞) curves revealed a 0.70±0.27 charge increase in the activation-gate valency (z m) following 8-Br-cAMP perfusion. Similar responses were observed when Ba2+ was the charge carrier, where z m was increased by 1.33±0.34 charges (n=8). The membrane potential for half activation (V 1/2) was also significantly shifted 6 mV more negative for I Ba (mean, n=8). The time course for I Ba (and I Ca) activation was well described by second-order m 2 kinetics. The derived time constant for activation (τm) was voltage-dependent, and the τm/V relation shifted negatively after 8-Br-cAMP treatment. Ca2+ channel gating rates were derived from the (τm) and m 2∞ values according to a Hodgkin-Huxley type m 2 activation process. The forward rate (α m) for channel activation was increased by 8-Br-cAMP at membrane potentials ≥0 mV, and the backward rate (βm) decreased at potentials ≤ +10 mV. Time-dependent inactivation of I Ca consisted of a slowly decaying component (τh ≈ 300 ms) and a “non-inactivating” steady-state component. The currents contributed by the two inactivation processes displayed different voltage dependences, the effects of 8-Br-cAMP being exclusively on the slowly inactivating L-type component.

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  1. Department of Physiology and Biophysics, University of Cincinnati College of Medicine, 231 Bethesda Avenue, 45267-0576, Cincinnati, Ohio, USA

    Craig A. Doupnik & Raymund Y. K. Pun

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  1. Craig A. Doupnik
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  2. Raymund Y. K. Pun
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Doupnik, C.A., Pun, R.Y.K. Cyclic AMP-dependent phosphorylation modifies the gating properties of L-type Ca2+ channels in bovine adrenal chromaffin cells. Pflugers Arch. 420, 61–71 (1992). https://doi.org/10.1007/BF00378642

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  • DOI: https://doi.org/10.1007/BF00378642

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