Moore R B, Manery J F, Still J, Mankad V N
Department of Pediatrics, University of South Alabama, Mobile 36617.
Biochem Cell Biol. 1989 Feb-Mar;67(2-3):137-46. doi: 10.1139/o89-021.
The activities of acetylcholinesterase and Ca2+ + Mg2+ ATPase were measured following treatment of human erythrocyte membranes with nonsolubilizing and solubilizing concentrations of Triton X-100. A concentration of 0.1% (v/v) Triton X-100 caused a significant inhibition of both enzymes. The inhibition appears to be caused by perturbations in the membrane induced by Triton X-100 incorporation. No acetylcholinesterase activity and little Ca2+ + Mg2+ ATPase activity were detected in the supernatant at 0.05% Triton X-100 although this same detergent concentration induced changes in the turbidity of the membrane suspension. Also, no inhibition of soluble acetylcholinesterase was observed over the entire detergent concentration range. The inhibition of these enzymes at 0.1% Triton X-100 was present over an eightfold range of membrane protein in the assay indicating an independence of the protein/detergent ratio. The losses in activities of these two enzymes could be prevented by either including phosphatidylserine in the Triton X-100 suspension or using Brij 96 which has the same polyoxyethylene polar head group but an oleyl hydrophobic tail instead of the p-tert-octylphenol group of Triton X-100. The results are discussed in regard to the differential recovery of enzyme activities over the entire detergent concentration range.
在用非增溶浓度和增溶浓度的 Triton X - 100 处理人红细胞膜后，测定了乙酰胆碱酯酶和 Ca2 + + Mg2 + ATP 酶的活性。0.1%（v/v）的 Triton X - 100 浓度会显著抑制这两种酶。这种抑制似乎是由 Triton X - 100 掺入诱导的膜扰动引起的。在 0.05% Triton X - 100 时，上清液中未检测到乙酰胆碱酯酶活性，Ca2 + + Mg2 + ATP 酶活性也很低，尽管相同的去污剂浓度会引起膜悬浮液浊度的变化。此外，在整个去污剂浓度范围内均未观察到可溶性乙酰胆碱酯酶受到抑制。在 0.1% Triton X - 100 时对这些酶的抑制在测定中膜蛋白的八倍范围内都存在，表明与蛋白质/去污剂比例无关。这两种酶活性的损失可以通过在 Triton X - 100 悬浮液中加入磷脂酰丝氨酸或使用 Brij 96 来防止，Brij 96 具有相同的聚氧乙烯极性头部基团，但疏水尾部是油基而不是 Triton X - 100 的对叔辛基苯酚基团。针对整个去污剂浓度范围内酶活性的差异恢复情况对结果进行了讨论。
