School of Molecular Bioscience, Kolling Institute, University of Sydney, Sydney, Australia.
Biophys J. 2013 Apr 16;104(8):1676-84. doi: 10.1016/j.bpj.2013.03.019.
We examined the response of Na(+),K(+)-ATPase (NKA) to monensin, a Na(+) ionophore, with and without ouabain, an NKA inhibitor, in suspensions of human erythrocytes (red blood cells). A combination of (13)C and (23)Na NMR methods allowed the recording of intra- and extracellular Na(+), and (13)C-labeled glucose time courses. The net influx of Na(+) and the consumption of glucose were measured with and without NKA inhibited by ouabain. A Bayesian analysis was used to determine probability distributions of the parameter values of a minimalist mathematical model of the kinetics involved, and then used to infer the rates of Na(+) transported and glucose consumed. It was estimated that the numerical relationship between the number of Na(+) ions transported by NKA per molecule of glucose consumed by a red blood cell was close to the ratio 6.0:1.0, agreeing with theoretical prediction.
我们研究了人红细胞悬液(红细胞)中钠钾-ATP 酶(NKA)对莫能菌素(一种钠离子载体)的反应,以及有无哇巴因(NKA 抑制剂)的情况下的反应。结合 (13)C 和 (23)Na NMR 方法,可记录细胞内外的 Na(+)和 (13)C 标记的葡萄糖时程。使用和不使用哇巴因抑制 NKA 的情况下,测量 Na(+)的净流入和葡萄糖的消耗。贝叶斯分析用于确定所涉及的动力学的最小数学模型的参数值的概率分布,然后用于推断 Na(+)的转运速率和葡萄糖的消耗速率。据估计,NKA 每转运一分子葡萄糖消耗的 Na(+)离子数与红细胞的数值关系接近 6.0:1.0 的比例,与理论预测相符。