Modulation of rat atrial G protein-coupled K+ channel function by phospholipids.

1. G protein-gated K+ channels (KACh channels) in the heart and brain are activated by the betagamma subunit of inhibitory G protein. Phosphatidylinositol-4,5-bisphosphate (PIP2) has recently been reported to directly activate KACh channels (GIRK) expressed in oocytes, as well as to support activation by the betagamma subunit in the presence of Na+. We examined the effect of Na+, PIP2 and other phospholipids on the KACh channel to understand better their role in KACh channel activation and modulation. 2. In atrial membrane patches, none of the phospholipids tested including PIP2 caused activation of the KACh channel in either the presence or the absence of 30 mM Na+. PIP2 (3 microM) and other phospholipids (30 microM) blocked acetylcholine-induced activation of the KACh channel. 3. When KACh channels were first activated with GTPgammaS, however, all phospholipids (100 microM) tested augmented the KACh channel activity 1.5- to 2-fold. Phosphatidylinositol-4-phosphate (PIP) and PIP2 were an order of magnitude more potent than other phospholipids. The increase in KACh channel activity was the result of a shift in the gating mode of the channel from a short-lived to a longer-lived open state. Such a modulatory effect was qualitatively similar to that produced by intracellular ATP. Trypsin blocked the ATP effect but not the phospholipid effect on the KACh channel kinetics. 4. The phosphate group linked to the glycerol backbone was important for KACh channel modulation by phospholipids. The higher potency of PIP and PIP2 was due to the presence of inositol phosphates. 5. Intracellular Na+ (30 mM) increased the frequency of KACh channel opening approximately 2-fold if the channels were already active, but did not affect modulation by phospholipids. The effects of Na+ and phospholipids on KACh channel activity were additive. 6. A low concentration of ATP (20 microM), which had no effect on the KACh channel by itself, potentiated the stimulatory action of phospholipids, indicating that ATP and phospholipids interacted to modulate KACh channel function. 7. We conclude that exogenously applied PIP2 and other phospholipids block agonist-mediated KACh channel activation. However, if the KACh channel is already activated with GTPgammaS, phospholipids augment the existing activity by increasing the number of longer-lived channel openings. The evidence for and against the role of PIP and PIP2 in the stimulatory effect of ATP on the KACh channel is presented and discussed.