Riccio P, Marra E, Passarella S, Quagliariello E
Boll Soc Ital Biol Sper. 1979 Dec 15;55(23):2412-8.
The binding of 3H-NQNO in submitochondrial particles was determined by measuring the radioactivity in the supernatants as well as in the sediments after centrifugation of particles suspensions containing different amounts of 3H-NQNO. From the binding data Scatchard plots were constructed showing a large amount of aspecific binding depending on the particles preparation and concentration. In the presence of saturating concentrations of either antimycin or unlabelled NQNO (2-n-Nonyl-4-hydroxy-quinolinee-N-oxide) that remove or prevent the specific binding of 3H-NQNO, it is possible to evaluate the aspecific component of 3H-NQNO binding and to subtracte it from the experimental binding data by graphyc correction according to (3). The straight line from the corrected points gives the specific binding parameters: number of specific binding sites: about 0,5 moles 3H-NQNO/ moles cytochrome b and KD= 50 nM.
通过测量含有不同量³H-NQNO的颗粒悬浮液离心后上清液和沉淀物中的放射性,来确定³H-NQNO在亚线粒体颗粒中的结合情况。根据结合数据构建Scatchard图,结果显示,取决于颗粒制备和浓度,存在大量非特异性结合。在存在饱和浓度的抗霉素或未标记的NQNO(2-正壬基-4-羟基喹啉-N-氧化物)的情况下,它们可去除或阻止³H-NQNO的特异性结合,根据(3)通过图形校正从实验结合数据中评估并减去³H-NQNO结合的非特异性成分成为可能。校正点的直线给出特异性结合参数:特异性结合位点数量:约0.5摩尔³H-NQNO/摩尔细胞色素b,KD = 50 nM。