Tirri R, Bowler K
Ann Acad Sci Fenn Biol. 1975(204):1-13.
In immature rats the temperature dependence of Na+-K+ ATPase of a crude homogenate of brain shows a compex curve with two activity maxima. When EDTA was present in the homogenization medium the curve obtained was of simpler, curvilinear type showing an increased activity at temperatures above 20 degrees C. The Na+-K+ ATPase activity in similar preparation from adult brain were not complex but curvilinear whether EDTA was used or not; however, EDTA increased the activity at temperatures above 20 degrees C. When such chelating agents as EDTA or histidine were used in preparation of microsomes from immature rat brain, the temperature dependence curve of Na+-K+ ATPase in this membrane fraction was changed to a steeper and simpler curve with increased activity especially at temperatures above 20 degrees. These agents, however, did not eliminate totally the complex shape of curve found in microsomes prepared without the presence of any chelating agents. When microsomes were prepared by using NaI-technique the temperature dependence of this enzyme was linear between temperatures 15-14 degrees, the activity being 4-5-fold higher than in the ordinary microsomal preparation. A stimulation of the Mg2+ ATPase by Na+ ions (100mM) was found at temperatures below 30 degrees but an inhibition by the same concentration of Na+ at upper temperatures. This effect together with the lowered activity due to bivalent metal ions (e.g. Ca2+, Cu2+) in 'crude' preparations was thought to be reasons for the complex shape of temperature dependence curve of Na+-K+ ATPase activity.
在未成熟大鼠中,脑粗匀浆中Na⁺-K⁺ATP酶的温度依赖性呈现出一条具有两个活性最大值的复杂曲线。当匀浆介质中存在乙二胺四乙酸(EDTA)时,所得到的曲线为较简单的曲线类型,在20℃以上温度时活性增加。来自成年大鼠脑的类似制剂中的Na⁺-K⁺ATP酶活性无论是否使用EDTA都不是复杂的而是曲线型的;然而,EDTA在20℃以上温度时增加了活性。当在从未成熟大鼠脑制备微粒体时使用诸如EDTA或组氨酸之类的螯合剂时,该膜部分中Na⁺-K⁺ATP酶的温度依赖性曲线变为更陡峭和更简单的曲线,活性增加,特别是在20℃以上温度时。然而,这些试剂并没有完全消除在没有任何螯合剂存在的情况下制备的微粒体中发现的复杂曲线形状。当使用碘化钠(NaI)技术制备微粒体时,该酶的温度依赖性在15 - 40℃之间呈线性,活性比普通微粒体制剂高4 - 5倍。在30℃以下温度时发现Na⁺离子(100mM)刺激Mg²⁺ATP酶,但在较高温度时相同浓度的Na⁺则产生抑制作用。这种效应以及由于“粗制”制剂中存在二价金属离子(例如Ca²⁺、Cu²⁺)而导致的活性降低被认为是Na⁺-K⁺ATP酶活性温度依赖性曲线形状复杂的原因。