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采用低渗溶血,高速旋离沉淀方法制得人红细胞膜制剂。测得Na~++K~+-ATP酶活力为6.15nmo P/min/mg蛋白质。测活反应系统中加入专一抑制剂乌本苷,一般可以用测活反应系统中缺K~+,Na~+来代替。在27～37℃范围内,Na~+-K~+-ATP酶对ATP的km值随温度上升而有增加。反应活化能为1.8×10~(-4)Cal/mol(卡/克分子)。 测活系统中加入Co~(2+),Ni~(2+),Zn~(2+),Cd~(2+),或Pb~(2+),对Na~+-K~+-ATP酶没有明显的影响,Sn~(2+)在10~(-4)M时有抑制Na~+-K~+-ATP酶的现象。Mn~(2+)能提高总ATP酶及Mg~(2+)-ATP酶活力,影响Na~+-K~+-ATP酶活力的测定。实验表明Ma~(2+)对乌本苷抑制Na~+-K~+-ATP酶的效应有拮抗作用。 没有发现胆汁酸盐对Na~+-K~+-ATP酶有明显的激活或抑制效应。
Using low permeability hemolysis, high speed spin-off precipitation method of human erythrocyte membrane preparation. The Na ~ + + K ~ + -ATPase activity was measured as 6.15nmo P / min / mg protein. Survival reaction system by adding a specific inhibitor of ouabain glycosides, the general reaction system can be used live lack of K ~ +, Na ~ + instead. In the range of 27 ~ 37 ℃, the ATP value of Na ~ + -K ~ + -ATPase increased with the increase of temperature. The activation energy is 1.8 × 10 -4 Cal / mol (cal / mol). The activity of Na ~ + - K ~ + in the presence of Co ~ (2 +), Ni ~ (2+), Zn ~ (2+), Cd 2+, ATPase had no obvious effect. Sn 2+ could inhibit Na ~ + -K ~ + -ATPase at 10 ~ (-4) M. Mn 2+ can increase the activity of total ATPase and Mg 2 + - ATPase, and affect the activity of Na ~ + -K ~ + -ATPase. Experiments show that Ma2 + has antagonistic effect on the inhibitory effect of ouabain on Na ~ + -K ~ + -ATPase. No bile salts were found to significantly activate or inhibit Na ~ + -K ~ + -ATPase.