Direct control of a large conductance K(+)-selective channel by G-proteins in adrenal chromaffin granule membranes.
We report here the presence of a Ca(2+)-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 alpha subunits for immunoblot analysis, we were able to identify the presence of the inhibitory G(i)2 and G(i)3 subtypes, as well as the stimulatory G(o) and Gs subtypes, but not G(i)1 in adrenal chromaffin granules. Furthermore, functional analysis of the K(+)-channel incorporated into planar lipid bilayers showed that GDP beta S and GTP gamma S 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.
Arispe
N
N
Laboratory of Cell Biology and Genetics, NIDDK, National Institutes of Health, Bethesda, MD 20892, USA.
De Mazancourt
P
P
Rojas
E
E
D000313:Adrenal Medulla / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002417:Cattle; D002837:Chromaffin Granules / Q000187:drug effects / Q000378:metabolism*; D004553:Electric Conductivity; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D008564:Membrane Potentials; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D011941:Receptors, Adrenergic / Q000378:metabolism; D012969:Sodium Fluoride / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology
D000906:Antibodies; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D011941:Receptors, Adrenergic; D013873:Thionucleotides; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C023427:guanosine 5'-O-(2-thiodiphosphate); D012969:Sodium Fluoride; D019204:GTP-Binding Proteins