红细胞膜中钠钾ATP酶测定的研究
Studies on the determination of Na-K-ATPase in red blood cell membranes.
作者信息
Eder K, Reichlmayr-Lais A M, Kirchgessner M
机构信息
Institut für Ernährungsphysiologie, Technische Universität München-Weihenstephan, Freising, F.R.G.
出版信息
J Trace Elem Electrolytes Health Dis. 1989 Sep;3(3):151-9.
The present paper is concerned with Na-K-ATPase. In the first part it deals with the effect of such factors as Na+, K+, Mg2+, ATP, pH on activity investigated using a soluble Na-K-ATPase from porcine cerebral cortex. In the second part a method for assaying red blood cell Na-K-ATPase is described. This method consists of specific steps for washing and preparation of red cell membranes, pretreatment of Na-K-ATPase with sodium dodecyl sulfate (SDS) and bovine serum albumin (BSA) and determination of enzymatic released inorganic phosphate. Na-K-ATPase prepared and treated in this way shows a higher activity than the conventionally prepared form of enzyme, as latent enzyme sites are exposed by the SDS treatment. The kinetic behaviour of Na-K-ATPase prepared and assayed by using this method is characterized and compared with that of soluble Na-K-ATPase.
本文关注钠钾ATP酶。第一部分探讨了诸如Na⁺、K⁺、Mg²⁺、ATP、pH等因素对使用猪脑皮层可溶性钠钾ATP酶所研究的活性的影响。第二部分描述了一种测定红细胞钠钾ATP酶的方法。该方法包括洗涤和制备红细胞膜的特定步骤、用十二烷基硫酸钠（SDS）和牛血清白蛋白（BSA）对钠钾ATP酶进行预处理以及测定酶促释放的无机磷酸盐。以这种方式制备和处理的钠钾ATP酶比传统制备的酶形式具有更高的活性，因为SDS处理暴露了潜在的酶位点。对使用该方法制备和测定的钠钾ATP酶的动力学行为进行了表征，并与可溶性钠钾ATP酶的动力学行为进行了比较。
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