Anner B M, Moosmayer M
J Biochem Biophys Methods. 1982 Feb;5(6):299-306. doi: 10.1016/0165-022x(82)90001-x.
A microprocedure for the preparation of Na,K-ATPase-containing liposomes with a minimal starting material (200 microgram) of purified Na,K-ATPase is presented. Phosphatidylcholine is added gradually to cholate-solubilized Na,K-ATPase of various concentrations and the lipid-induced decrease in enzyme activity is monitored. After removal of the detergent by dialysis, the transport parameters of the resulting Na,K-ATPase-liposomes are established by a microassay. By relating the transport properties to the Na,K-ATPase activity preset before dialysis, a procedure is developed which allows to prepare standardized Na,K-ATPase-liposomes with predictable transport properties.
本文介绍了一种微量制备含钠钾ATP酶脂质体的方法,该方法所需的纯化钠钾ATP酶起始材料最少(200微克)。将磷脂酰胆碱逐渐添加到不同浓度的胆酸盐增溶的钠钾ATP酶中,并监测脂质诱导的酶活性降低。通过透析去除去污剂后,通过微量测定法确定所得钠钾ATP酶脂质体的转运参数。通过将转运特性与透析前预设的钠钾ATP酶活性相关联,开发了一种程序,该程序允许制备具有可预测转运特性的标准化钠钾ATP酶脂质体。