title: Cytoplasmic unsaturated free fatty acids inhibit ATP-dependent gating of the G protein-gated K(+) channel.
abstract: This study reports the identification of an endogenous inhibitor of the G protein-gated (K(ACh)) channel and its effect on the K(ACh) channel kinetics. In the presence of acetylcholine in the pipette, K(ACh) channels in inside-out atrial patches were activated by applying GTP to the cytoplasmic side of the membrane. In these patches, addition of physiological concentration of intracellular ATP (4 mM) upregulated K(ACh) channel activity approximately fivefold and induced long-lived openings. However, such ATP-dependent gating is normally not observed in cell-attached patches, indicating that an endogenous substance that inhibits the ATP effect is present in the cell. We searched for such an inhibitor in the cell. ATP-dependent gating of the K(ACh) channel was inhibited by the addition of the cytosolic fraction of rat atrial or brain tissues. The lipid component of the cytosolic fraction was found to contain the inhibitory activity. To identify the lipid inhibitor, we tested the effect of approximately 40 different lipid molecules. Among the lipids tested, only unsaturated free fatty acids such as oleic, linoleic, and arachidonic acids (0.2-2 microM) reversibly inhibited the ATP-dependent gating of native K(ACh) channels in atrial cells and hippocampal neurons, and of recombinant K(ACh) channels (GIRK1/4 and GIRK1/2) expressed in oocytes. Unsaturated free fatty acids also inhibited phosphatidylinositol-4, 5-bisphosphate (PIP(2))-induced changes in K(ACh) channel kinetics but were ineffective against ATP-activated background K(1) channels and PIP(2)-activated K(ATP) channels. These results show that during agonist-induced activation, unsaturated free fatty acids in the cytoplasm help to keep the cardiac and neuronal K(ACh) channels downregulated by antagonizing their ATP-dependent gating. The opposing effects of ATP and free fatty acids represent a novel regulatory mechanism for the G protein-gated K(+) channel.
authors.affiliation: Department of Physiology, Finch University of Health Sciences, The Chicago Medical School, North Chicago, Illinois 60064, USA. kimd@mis.finchcms.edu
authors.forename: D
authors.initials: D
authors.lastname: Kim
authors.forename: A
authors.initials: A
authors.lastname: Pleumsamran
mesh_terms: D000109:Acetylcholine / Q000493:pharmacokinetics; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D016718:Arachidonic Acid / Q000032:analysis / Q000494:pharmacology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000494:pharmacology*; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006325:Heart Atria / Q000737:chemistry / Q000166:cytology; D006339:Heart Rate / Q000502:physiology; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology; D007700:Kinetics; D019787:Linoleic Acid / Q000032:analysis / Q000494:pharmacology; D009206:Myocardium / Q000737:chemistry / Q000166:cytology; D019301:Oleic Acid / Q000032:analysis / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D011188:Potassium / Q000493:pharmacokinetics; D015221:Potassium Channels / Q000502:physiology*; D024661:Potassium Channels, Inwardly Rectifying; D051381:Rats; D009435:Synaptic Transmission / Q000502:physiology
chemical_list: D005231:Fatty Acids, Unsaturated; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D019269:Phosphatidylinositol 4,5-Diphosphate; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D016718:Arachidonic Acid; D019301:Oleic Acid; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D019787:Linoleic Acid; D000109:Acetylcholine; D011188:Potassium