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培养的心肌细胞和神经母细胞瘤细胞对钠依赖性钙摄取的动力学研究。

Kinetic studies on sodium-dependent calcium uptake by myocardial cells and neuroblastoma cells in culture.

作者信息

Wakabayashi S, Goshima K

出版信息

Biochim Biophys Acta. 1981 Mar 20;642(1):158-72. doi: 10.1016/0005-2736(81)90146-2.

DOI:10.1016/0005-2736(81)90146-2
PMID:6784764
Abstract

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.

摘要

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。

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