Regulation of atrial muscarinic K+ channel activity by a cytosolic protein via G protein-independent pathway.
Rapid desensitization of the muscarinic K+ current (KACh current) is observed in cell-attached patches with 10 microM acetylcholine in the pipette. When inside-out patches were formed within approximately 1 s after formation of cell-attached patches and GTP was applied to the cytoplasmic side of the membrane, desensitization was not observed, indicating that a cytosolic factor mediated the desensitization. Applying the atrial cytosolic extract directly to the cytoplasmic side of such inside-out patches elicited a rapid desensitization of the KACh current. ATP (1-4 mM) reversed this effect of the cytosol and reverted the KACh channel to the undesensitized state. These effects of ATP and cytosol on the KACh channel could occur in the absence of GTP or in the presence of 100 microM guanosine 5'-O-(3-thiotriphosphate), indicating that G protein was not involved. Treatment of the cytosol with proteases (trypsin, chymotrypsin, bacterial protease) or heat denaturation abolished the effect of the cytosol on the KACh channel kinetics, indicating that the cytosolic factor was a protein. Functional assay of the fractions collected from gel filtration column indicated that the molecular mass of the native protein was 95-130 kDa. We conclude that a large cytosolic protein mediates the rapid desensitization of the KACh channel current via a G protein-independent pathway.
Hong
S G
SG
Department of Physiology and Biophysics, Finch University of Health Sciences, Chicago Medical School, North Chicago, Illinois 60064, USA.
Pleumsamran
A
A
Kim
D
D
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002457:Cell Extracts / Q000494:pharmacology; D003600:Cytosol / Q000737:chemistry / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000502:physiology*; D006325:Heart Atria; D007700:Kinetics; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D011506:Proteins / Q000502:physiology; D051381:Rats
D002457:Cell Extracts; D015221:Potassium Channels; D047428:Protein Kinase Inhibitors; D011506:Proteins; D000255:Adenosine Triphosphate; D010744:Phosphoric Monoester Hydrolases; D019204:GTP-Binding Proteins