Kim Y K, Cho H J, Kim W T, Cho K S
Department of Agricultural Chemistry, Chungbuk National University, Cheongju, Korea.
Biochem Biophys Res Commun. 1997 Jan 13;230(2):247-50. doi: 10.1006/bbrc.1996.5936.
Major parts of microsomes prepared from the epithelial cells of porcine trachea were tight-sealed vesicles since they showed a saturation of 45Ca2+ uptake and spontaneous releases of stored 45Ca2+ by the treatments of Ca2+-ionophore and Ca2+ channel agonists. In the presence of caffeine (10 mM), the maximal release of microsomal 45Ca2+ was observed at the extramicrosomal Ca2+ concentrations between 0.1 approximately 1 microM and at below or above this range of Ca2+ concentration the releases were decreased, forming a bell-shaped curve. These results indicate that the microsomal 45Ca2+ releases were mediated by ryanodine receptor, a caffeine-sensitive Ca2+ channel. Caffeine (10 mM) released 30.2 +/- 5.9% of microsomal 45Ca2+ while inositol 1,4,5-trisphosphate (InsP3, 10 microM) released 18.4 +/- 3.0% of the stored 45Ca2+. Caffeine-induced and InsP3-induced 45Ca2+ releases were additive, implying that these two types of 45Ca2+ releases are from physically distinct microsomes. Procaine, an antagonist of ryanodine receptor, selectively blocked the effect of caffeine but not the effect of InsP3. The results suggest that the epithelial cells of porcine trachea have caffeine-sensitive Ca2+ store in addition to InsP3-sensitive one.
从猪气管上皮细胞制备的微粒体的主要部分是紧密密封的囊泡,因为它们表现出45Ca2+摄取的饱和以及通过钙离子载体和钙通道激动剂处理后储存的45Ca2+的自发释放。在咖啡因(10 mM)存在下,在微粒体外钙浓度介于0.1至约1 microM之间时观察到微粒体45Ca2+的最大释放,而在该钙浓度范围以下或以上时释放减少,形成钟形曲线。这些结果表明微粒体45Ca2+的释放是由兰尼碱受体介导的,兰尼碱受体是一种对咖啡因敏感的钙通道。咖啡因(10 mM)释放了微粒体45Ca2+的30.2±5.9%,而肌醇1,4,5-三磷酸(InsP3,10 microM)释放了储存的45Ca2+的18.4±3.0%。咖啡因诱导的和InsP3诱导的45Ca2+释放是相加的,这意味着这两种类型的45Ca2+释放来自物理上不同的微粒体。普鲁卡因是兰尼碱受体的拮抗剂,它选择性地阻断了咖啡因的作用,但不阻断InsP3的作用。结果表明猪气管上皮细胞除了有对InsP3敏感的钙储存外,还有对咖啡因敏感的钙储存。