Lubberding H J, Zimmer G, van Walraven H S, Schrickx J, Kraayenhof R
Eur J Biochem. 1983 Dec 1;137(1-2):95-9. doi: 10.1111/j.1432-1033.1983.tb07800.x.
The ATPase complex is isolated and purified from membrane vesicles of the thermophilic cyanobacterium Synechococcus 6716 by octyl glucoside and cholic acid by a modification of the procedure for its extraction from spinach chloroplasts. The complex is purified by differential centrifugation and ammonium sulfate precipitation and by gel filtration on Sepharose 6B. The purified fraction, without any phycocyanin contamination, shows ATP hydrolysis activity and Pi/ATP exchange activity of 1564 and 350 nmol X min-1 X mg protein-1, respectively. N,N'-Dicyclohexylcarbodiimide inhibits the ATP hydrolysis activity of this purified fraction. On polyacrylamide gels most typical F1 ATPase polypeptides are identified, but the low-molecular weight polypeptides visible cannot be ascribed to the F0 part of the complex with certainty; non-identified bands around 30 kDa are also present.
通过对从菠菜叶绿体中提取该复合物的方法进行改进，利用辛基葡糖苷和胆酸从嗜热蓝藻集胞藻6716的膜囊泡中分离并纯化ATP酶复合物。通过差速离心、硫酸铵沉淀以及在琼脂糖6B上进行凝胶过滤对该复合物进行纯化。纯化后的组分无任何藻蓝蛋白污染，其ATP水解活性和Pi/ATP交换活性分别为1564和350 nmol·min⁻¹·mg蛋白⁻¹。N,N'-二环己基碳二亚胺抑制该纯化组分的ATP水解活性。在聚丙烯酰胺凝胶上鉴定出了大多数典型的F1 ATP酶多肽，但可见的低分子量多肽不能确定地归属于该复合物的F0部分；还存在30 kDa左右未鉴定的条带。
