Activation of the cloned muscarinic potassium channel by G protein beta gamma subunits.
Acetylcholine released during parasympathetic stimulation of the vagal nerve slows the heart rate through the activation of muscarinic receptors and subsequent opening of an inwardly rectifying potassium channel. The activation of these muscarinic potassium channels is mediated by a pertussis toxin-sensitive heterotrimeric GTP-binding protein (G protein). It has not been resolved whether exogenously applied G alpha or G beta gamma, or both, activate the channel. Using a heterologous expression system, we have tested the ability of different G protein subunits to activate the cloned muscarinic potassium channel, GIRK1. We report here that coexpression of GIRK1 with G beta gamma but not G alpha beta gamma in Xenopus oocytes results in channel activity that persists in the absence of cytoplasmic GTP. This activity is reduced by fusion proteins of the beta-adrenergic receptor kinase and of recombinant G alpha i-GDP, both of which are known to interact with G beta gamma. Moreover, application of recombinant G beta gamma, but not G alpha i-GTP-gamma S, activates GIRK1 channels. Thus G beta gamma appears to be sufficient for the activation of GIRK1 muscarinic potassium channels.
Howard Hughes Medical Institute, University of California, San Francisco 94143.
E
E
Reuveny
P A
PA
Slesinger
J
J
Inglese
J M
JM
Morales
J A
JA
Iñiguez-Lluhi
R J
RJ
Lefkowitz
H R
HR
Bourne
Y N
YN
Jan
L Y
LY
Jan
D000595:Amino Acid Sequence; D000818:Animals; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D019204:GTP-Binding Proteins / Q000502:physiology*; D006150:Guanine Nucleotides / Q000378:metabolism; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000502:physiology; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010446:Peptide Fragments / Q000502:physiology; D015221:Potassium Channels / Q000502:physiology*; D024661:Potassium Channels, Inwardly Rectifying; D011976:Receptors, Muscarinic / Q000502:physiology*; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014981:Xenopus
D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D006150:Guanine Nucleotides; D008565:Membrane Proteins; D010446:Peptide Fragments; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D011976:Receptors, Muscarinic; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D019204:GTP-Binding Proteins