Bender Kirsten, Wellner-Kienitz Marie-Cécile, Pott Lutz
Department of Physiology, Ruhr-University Bochum, D-4480 Bochum, Germany.
FEBS Lett. 2002 Oct 9;529(2-3):356-60. doi: 10.1016/s0014-5793(02)03426-9.
GIRK (G protein-activated inward-rectifying K(+) channel) channels, important regulators of membrane excitability in the heart and in the central nervous, are activated by interaction with betagamma subunits from heterotrimeric G proteins upon receptor stimulation. For activation interaction of the channel with phosphatidylinositol 4,5-bisphosphate (PtIns(4,5)P(2)) is conditional. Previous studies have provided evidence that in myocytes PtIns(4,5)P(2) levels relevant to GIRK channel regulation are under regulatory control of receptors activating phospholipase C. In the present study a phosphatidyl-4-phosphate 5-kinase was expressed in atrial myocytes by transient transfection. This did not affect basal properties of GIRK current activated by acetylcholine via M(2) receptors but completely abolished inhibition of guanosine triphosphate-gamma-S activated current by endothelin-1 or alpha-adrenergic agonists. We conclude that though PtIns(4,5)P(2) is conditional for channel gating, its normal level in the membrane is not limiting basal function of GIRK channels. Moreover, our data provide further evidence for a regulation of GIRK channels by alpha(1A) receptors and endothelin-A receptors, endogenously expressed in atrial myocytes, via depletion of PtIns(4,5)P(2).
GIRK(G蛋白激活的内向整流钾通道)通道是心脏和中枢神经系统中膜兴奋性的重要调节因子,在受体刺激时,通过与异源三聚体G蛋白的βγ亚基相互作用而被激活。通道与磷脂酰肌醇4,5-二磷酸(PtIns(4,5)P(2))的相互作用对于激活来说是有条件的。先前的研究已提供证据表明,在心肌细胞中,与GIRK通道调节相关的PtIns(4,5)P(2)水平受激活磷脂酶C的受体的调节控制。在本研究中,通过瞬时转染在心房肌细胞中表达了一种磷脂酰-4-磷酸5-激酶。这并不影响通过M(2)受体由乙酰胆碱激活的GIRK电流的基础特性,但完全消除了内皮素-1或α-肾上腺素能激动剂对鸟苷三磷酸-γ-S激活电流的抑制作用。我们得出结论,虽然PtIns(4,5)P(2)对于通道门控是有条件的,但其在膜中的正常水平并不限制GIRK通道的基础功能。此外,我们的数据为心房肌细胞中内源性表达的α(1A)受体和内皮素-A受体通过耗尽PtIns(4,5)P(2)对GIRK通道进行调节提供了进一步的证据。