Wagner J A, Cozens A L, Schulman H, Gruenert D C, Stryer L, Gardner P
Department of Medicine, Falk CVRC, Stanford University School of Medicine, California 94305.
Nature. 1991 Feb 28;349(6312):793-6. doi: 10.1038/349793a0.
Cystic fibrosis is associated with defective regulation of apical membrane chloride channels in airway epithelial cells. These channels in normal cells are activated by cyclic AMP-dependent protein kinase and protein kinase C. In cystic fibrosis these kinases fail to activate otherwise normal Cl- channels. But Cl- flux in cystic fibrosis cells, as in normal cells, can be activated by raising intracellular Ca2+ (refs 5-10). We report here whole-cell patch clamp studies of normal and cystic fibrosis-derived airway epithelial cells showing that Cl- channel activation by Ca2+ is mediated by multifunctional Ca2+/calmodulin-dependent protein kinase. We find that intracellular application of activated kinase and ATP activates a Cl- current similar to that activated by a Ca2+ ionophore, that peptide inhibitors of either the kinase or calmodulin block Ca2(+)-dependent activation of Cl- channels, and that a peptide inhibitor of protein kinase C does not block Ca2(+)-dependent activation. Ca2+/calmodulin activation of Cl- channels presents a pathway with therapeutic potential for circumventing defective regulation of Cl- channels in cystic fibrosis.
囊性纤维化与气道上皮细胞顶端膜氯化物通道的调节缺陷有关。正常细胞中的这些通道由环磷酸腺苷依赖性蛋白激酶和蛋白激酶C激活。在囊性纤维化中,这些激酶无法激活原本正常的氯离子通道。但是,与正常细胞一样,囊性纤维化细胞中的氯通量可以通过提高细胞内钙离子浓度来激活(参考文献5 - 10)。我们在此报告对正常和源自囊性纤维化的气道上皮细胞进行的全细胞膜片钳研究,结果表明钙离子对氯离子通道的激活是由多功能钙调蛋白依赖性蛋白激酶介导的。我们发现,在细胞内应用活化的激酶和三磷酸腺苷可激活一种类似于由钙离子载体激活的氯电流,该激酶或钙调蛋白的肽类抑制剂可阻断氯离子通道的钙离子依赖性激活,而蛋白激酶C的肽类抑制剂则不会阻断钙离子依赖性激活。钙离子/钙调蛋白对氯离子通道的激活提供了一条具有治疗潜力的途径,可绕过囊性纤维化中氯离子通道的调节缺陷。