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Direct control of a large conductance K+-selective channel by G-proteins in adrenal chromaffin granule membranes

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Abstract

We report here the presence of a Ca2+-independent K+-channel of large conductance in adrenal chromaffin cell secretory vesicle membranes which is controlled by inhibitory as well as stimulatory heterotrimeric GTP-binding proteins. Using antibodies against specific α subunits for immunoblot analysis, we were able to identify the presence of the inhibitory Gi2 and Gi3 subtypes, as well as the stimulatory G 0 and G s subtypes, but not Gi1 in adrenal chromaffin granules. Furthermore, functional analysis of the K+-channel incorporated into planar lipid bilayers showed that GDPβS and GTPγS have opposite effects on channel activity inducing interconversions between a low and a high open-probability state. Consistent with these findings, the same antibodies antagonized the effects of the nonhydrolyzable analogues on the open probability of the K+-channel.

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Additional information

The authors thank Dr. H.B. Pollard for many invaluable comments and discussions on the physiology of chromaffin secretory vesicles; Drs. A.M. Spiegel and P.K. Goldsmith for providing the antibodies used in this work, and Dr. G. Lee for providing us with a highly purified preparation of chromaffin granules. Thanks are also given to David Mears for revising the manuscript, and to B. Chidakel for electronic design and construction.

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Arispe, N., De Mazancourt, P. & Rojas, E. Direct control of a large conductance K+-selective channel by G-proteins in adrenal chromaffin granule membranes. J. Membarin Biol. 147, 109–119 (1995). https://doi.org/10.1007/BF00233540

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Key words

  • K+ channel
  • G-protein
  • Secretory vesicles
  • Antibodies
  • GTPγS
  • GDPβS