Lüdi H, Hasselbach W
J Chromatogr. 1984 Aug 3;297:111-7. doi: 10.1016/s0021-9673(01)89034-8.
The state of aggregation of purified sarcoplasmic reticulum adenosine triphosphatase (ATPase) was investigated by high-performance liquid chromatography (LKB TSK-G 4000 SW column) in the presence of various detergents: sodium dodecylsulphate, dodecyl octaethylene glycol monoether (C12E8), sodium deoxycholate, Triton X-100 and myristoylglycerophosphocholine. When the protein (5 mg ml-1) was solubilized with detergent (2 mg per mg protein) and the column was equilibrated with 1 mg ml-1 of the respective detergent, a molecular weight for the monomeric ATPase protein ranging from 100,000 to 200,000 was obtained. In addition to the monomeric form, significant amounts (more than 20%) of aggregated ATPase protein were observed when C12E8 or deoxycholate was used. These results are in agreement with the observation of a great tendency for self-aggregation of the ATPase protein in conventional gel filtration chromatography and ultracentrifugation experiments. The dimeric form of the ATPase protein was detected only when deoxycholate and, probably, when C12E8 was used.
在各种去污剂存在的情况下，通过高效液相色谱法（LKB TSK - G 4000 SW柱）研究了纯化的肌浆网腺苷三磷酸酶（ATPase）的聚集状态，这些去污剂包括：十二烷基硫酸钠、十二烷基八乙二醇单醚（C12E8）、脱氧胆酸钠、Triton X - 100和肉豆蔻酰甘油磷酸胆碱。当蛋白质（5 mg/ml）用去污剂（每毫克蛋白质2 mg）溶解，且柱子用1 mg/ml各自的去污剂平衡时，得到的单体ATPase蛋白的分子量在100,000至200,000之间。除了单体形式外，当使用C12E8或脱氧胆酸钠时，观察到大量（超过20%）聚集的ATPase蛋白。这些结果与在传统凝胶过滤色谱法和超速离心实验中观察到的ATPase蛋白高度的自我聚集倾向一致。仅当使用脱氧胆酸钠以及可能使用C12E8时，检测到了ATPase蛋白的二聚体形式。
