Ward J, Cardoso de Almeida M L, Turner M J, Etges R, Bordier C
Mol Biochem Parasitol. 1987 Feb;23(1):1-7. doi: 10.1016/0166-6851(87)90180-0.
The technique of phase separation in a solution of the non-ionic detergent Triton X-114 was used to measure the enzymatic conversion of a membrane protein to a soluble product via removal of a hydrophobic moiety. The substrate was the major surface protein (p63), of Leishmania promastigotes and the enzyme was a phospholipase C purified from Trypanosoma brucei. This membrane-bound enzyme is responsible for the cleavage of the hydrophobic lipid membrane anchor of the variant surface glycoprotein (VSG), of T. brucei. The assay is fast, simple and uses small amounts of reagents. It has been used to determine the pH optimum, thermal resistance, and the sensitivity to inhibitors of the trypanosomal phospholipase.
利用非离子去污剂Triton X-114溶液中的相分离技术，通过去除疏水部分来测量膜蛋白向可溶性产物的酶促转化。底物是利什曼原虫前鞭毛体的主要表面蛋白（p63），酶是从布氏锥虫纯化的磷脂酶C。这种膜结合酶负责切割布氏锥虫可变表面糖蛋白（VSG）的疏水脂质膜锚定物。该测定方法快速、简单，且使用少量试剂。它已被用于确定锥虫磷脂酶的最适pH值、耐热性以及对抑制剂的敏感性。
