Dascal N, Doupnik C A, Ivanina T, Bausch S, Wang W, Lin C, Garvey J, Chavkin C, Lester H A, Davidson N
Department of Physiology and Pharmacology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6758-62. doi: 10.1073/pnas.92.15.6758.
Coexpression in Xenopus oocytes of the inwardly rectifying guanine nucleotide binding (G)-protein-gated K channel GIRK1 with a myristoylated modification of the (putative) cytosolic C-terminal tail [GIRK1 aa 183-501 fused in-frame to aa 1-15 of p60src and denoted src+ (183-501)] leads to a high degree of inhibition of the inward G-protein-gated K+ current. The nonmyristoylated segment, src- (183-501), is not active. Although some interference with assembly is not precluded, the evidence indicates that the main mechanism of inhibition is interference with functional activation of the channel by G proteins. In part, the tail functions as a blocking particle similar to a "Shaker ball"; it may also function by competing for the available supply of free G beta gamma liberated by hormone activation of a seven-helix receptor. The non-G-protein-gated weak inward rectifier ROMK1 is less effectively inhibited, and a Shaker K channel was not inhibited. Immunological assays show the presence of a high concentration of src+ (183-501) in the plasma membrane and the absence of any membrane forms for the nonmyristoylated segment.
内向整流型鸟嘌呤核苷酸结合(G)蛋白门控钾通道GIRK1与(假定的)胞质C末端尾巴经肉豆蔻酰化修饰(GIRK1的183 - 501位氨基酸与p60src的1 - 15位氨基酸读框内融合并命名为src + (183 - 501))在非洲爪蟾卵母细胞中共表达,会导致内向G蛋白门控钾电流受到高度抑制。非肉豆蔻酰化片段src - (183 - 501)无活性。尽管不排除对组装有一些干扰,但证据表明抑制的主要机制是干扰G蛋白对通道的功能激活。该尾巴部分起到类似于“Shaker球”的阻断颗粒的作用;它也可能通过竞争七螺旋受体激素激活所释放的游离Gβγ的可用供应来发挥作用。非G蛋白门控的弱内向整流器ROMK1受到的抑制效果较差,而Shaker钾通道未受抑制。免疫分析显示质膜中存在高浓度的src + (183 - 501),且非肉豆蔻酰化片段不存在任何膜形式。