Dupuis A, Duszynski J, Vignais P V
Biochem Biophys Res Commun. 1987 Jan 15;142(1):31-7. doi: 10.1016/0006-291x(87)90447-5.
The fluorescent probe, 6-acryloyl-2-dimethylaminonaphtalene (acrylodan) was reacted with the oligomycin-sensitivity conferring protein (OSCP). Acrylodan bound covalently to the single cysteinyl residue of the protein. Acrylodan-OSCP was fully competent in conferring oligomycin sensitivity to the mitochondrial F0-F1 ATPase complex. The fluorescence emission peak of acrylodan-OSCP was blue-shifted compared to that of an acrylodan-mercaptoethanol adduct, which means that acrylodan experiences a hydrophobic environment in OSCP. Binding of acrylodan-OSCP to the isolated F1 was accompanied by a red shift of fluorescence. It was achieved in less than 1 s at 25 degrees C. The titration curve revealed one high affinity OSCP binding site per F1. Acrylodan-OSCP appears to be an interesting tool for studying the dynamics of structural changes within the mitochondrial ATPase complex.
荧光探针6-丙烯酰基-2-二甲基氨基萘（丙烯罗丹）与寡霉素敏感性赋予蛋白（OSCP）发生反应。丙烯罗丹与该蛋白的单个半胱氨酸残基共价结合。丙烯罗丹标记的OSCP能够完全赋予线粒体F0-F1 ATP酶复合体寡霉素敏感性。与丙烯罗丹-巯基乙醇加合物相比，丙烯罗丹标记的OSCP的荧光发射峰发生了蓝移，这意味着丙烯罗丹在OSCP中处于疏水环境。丙烯罗丹标记的OSCP与分离出的F1结合伴随着荧光的红移。在25℃下不到1秒即可完成。滴定曲线显示每个F1有一个高亲和力的OSCP结合位点。丙烯罗丹标记的OSCP似乎是研究线粒体ATP酶复合体内结构变化动力学的一个有趣工具。
