Wakabayashi S, Goshima K
Biochim Biophys Acta. 1981 Mar 20;642(1):158-72. doi: 10.1016/0005-2736(81)90146-2.
Kinetic analyses were made on intracellular Na+-dependent Ca2+ uptake by myocardial cells and neuroblastoma cells (N-18 strain) in culture. Cells loaded with various concentrations of Na+ could be prepared by incubating them in Ca2+-free medium containing various concentrations of Na+. Cells pre-loaded with various concentrations of Na+ were incubated in medium containing Ca2+ and 45Ca. The resulting 45Ca uptake by the two types of cell depended greatly on the initial intracellular concentrations of Na+. Lineweaver-Burk plots of the initial rate of Ca2+ uptake against the external concentration of Ca2+ fitted well to straight lines obtained by linear regression (r greater than 0.95). This result shows that Ca2+ uptake by the two types of cell was achieved by a carrier-mediated transport system. This Na+-dependent Ca2+ uptake was accompanied by Na+ release and the ratio of Na+ release to Ca2+ uptake was close to 3 : 1. A comparison of the kinetic data between myocardial cells and N-18 cells suggested that N-18 cells possess a carrier showing the same properties as that of myocardial cells, i.e.: (1) a similar dependency on the intracellular concentration of Na+; (2) the coincidence of the apparent Michaelis constants for Ca2+ (0.1 mM); (3) the similarities of the Ki values for Co2+, Sr2+ and Mg2+ (Co2+ less than Sr2+ less than Mg2+) and (4) a similar dependency on pH. However, the maximal initial rate, V, of N-18 cells was about 1/100 that of myocardial cells. The rate of Na+-dependent Ca2+ uptake by non-excitable cells was much lower than that by myocardial cells.
对培养的心肌细胞和神经母细胞瘤细胞(N - 18株)内依赖钠的钙离子摄取进行了动力学分析。通过将细胞在含有不同浓度钠的无钙培养基中孵育,可以制备出加载有不同浓度钠的细胞。将预先加载有不同浓度钠的细胞在含有钙离子和45钙的培养基中孵育。两种类型细胞对45钙的摄取很大程度上取决于细胞内钠的初始浓度。以钙离子摄取的初始速率对细胞外钙离子浓度所作的Lineweaver - Burk图很好地拟合了通过线性回归得到的直线(r大于0.95)。这一结果表明,两种类型细胞对钙离子的摄取是通过载体介导的转运系统实现的。这种依赖钠的钙离子摄取伴随着钠的释放,并且钠释放与钙离子摄取的比率接近3∶1。心肌细胞和N - 18细胞之间动力学数据的比较表明,N - 18细胞具有一种与心肌细胞性质相同的载体,即:(1)对细胞内钠浓度有相似的依赖性;(2)钙离子的表观米氏常数一致(0.1 mM);(3)钴离子、锶离子和镁离子的抑制常数Ki值相似(钴离子<锶离子<镁离子);(4)对pH有相似的依赖性。然而,N - 18细胞的最大初始速率V约为心肌细胞的1/100。非兴奋性细胞依赖钠的钙离子摄取速率远低于心肌细胞。