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Slow inhibition of N-type calcium channels with GTPγS reflects the basal G protein-GDP turnover rate

  • Cellular Neurophysiology
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Abstract

The inhibition of N-type Ca channels via a G protein pathway is a rapid mechanism for modulating Ca influx. It has been noted, however, that when G proteins are activated by guanosine 5′-O-(3-thiotriphosphate) (GTPγS), the speed of inhibition is greatly reduced, despite the pathway having fewer molecular steps. We explored this anomaly in chick dorsal root ganglion neurons by comparing Ca current inhibition using GTPγS with application of the G protein receptor agonist noradrenaline. Noradrenaline caused rapid Ca channel inhibition (τ~5 s), contrasting greatly with the ~70-fold slower rate observed with GTPγS. Additionally, the slow rate with GTPγS could be accelerated to near agonist-induced rates by application of noradrenaline, demonstrating that the inhibition with GTPγS was not perfusion limited and that the rate-limiting step was upstream from GTPγS binding. Our results suggest that in the absence of noradrenaline, G protein activation by GTPγS is impeded by the slow resting turnover of GDP/GTP. The rate at which inhibition develops with GTPγS (τ~350 s) is thus a direct and sensitive measure of resting GDP turnover.

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Acknowledgements

We are grateful for the technical assistance of Dr. Qi Li and to discussions with Luigi Gentile, Drs Terry Morris, Lyanne Schlichter and Jane Mitchell. This work was supported by Canadian Institutes for Health Research grant awards MOP38091 and MOP57716 and a Canada Research Chair to EFS.

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Authors and Affiliations

  1. Division of Cellular and Molecular Biology, Toronto Western Research Institute, University Health Network, Toronto, Ontario, M5T 2S8, Canada

    Allen W. Chan & Elise F. Stanley

  2. Department of Physiology, University of Toronto, Toronto, Ontario, M5S 1A8, Canada

    Allen W. Chan & Elise F. Stanley

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  1. Allen W. Chan
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  2. Elise F. Stanley
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Correspondence to Elise F. Stanley.

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Chan, A.W., Stanley, E.F. Slow inhibition of N-type calcium channels with GTPγS reflects the basal G protein-GDP turnover rate. Pflugers Arch - Eur J Physiol 446, 183–188 (2003). https://doi.org/10.1007/s00424-003-1030-2

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  • DOI: https://doi.org/10.1007/s00424-003-1030-2

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