Siegel G J, Tormay A, Candia O A
Biochim Biophys Acta. 1975 May 21;389(3):547-66.
A method is described for the extraction of microsomal ouabain-sensitive (a- + K+)-activated ATPase from separated frog skin epithelium. The method yields a microsomal fraction containing (Na+ K+)-stimulated activity in the range of 30- 40 nmol - mg -1 - min -1 at 26 degrees C. This portion which is also ouabain sensitive, is about half of the total activity in media containing Mg2+, Na+ and K+. These preparations also contain Mg2+-dependent or Ca2+-dependent activities which are not additive and which are not significantly affected by ouabain, Na+, K+ or Li+. The activations of the ouabain-sensitive ATPase activity by Mg2+, Na+, and K+ are similar to those described in other tissues. It is found that Li+ does not substitute for Na+ as an activator but in high concentrations does produce partial activation in the presence of Na+ with no K+. These results are pertinent to the reported observations of ouabain-sensitive Li+ flux across frog skin. It is concluded that this flux is not apparently due to a direct activating effect of Li+ on the sodium pump.
本文描述了一种从分离的蛙皮上皮中提取微粒体哇巴因敏感的(钠+钾)激活ATP酶的方法。该方法得到的微粒体部分在26℃时具有30 - 40 nmol·mg⁻¹·min⁻¹范围内的(钠+钾)刺激活性。这部分活性对哇巴因也敏感,约为含有镁离子、钠离子和钾离子的培养基中总活性的一半。这些制剂还含有镁离子依赖性或钙离子依赖性活性,它们不具有加和性,且不受哇巴因、钠离子、钾离子或锂离子的显著影响。镁离子、钠离子和钾离子对哇巴因敏感的ATP酶活性的激活作用与其他组织中描述的相似。发现锂离子不能替代钠离子作为激活剂,但在高浓度下,在无钾离子存在的情况下,与钠离子一起确实会产生部分激活作用。这些结果与报道的关于哇巴因敏感的锂离子跨蛙皮通量的观察结果相关。得出的结论是,这种通量显然不是由于锂离子对钠泵的直接激活作用。