Gibrat R, Romieu C, Grignon C
Biochim Biophys Acta. 1983 Dec 21;736(2):196-202. doi: 10.1016/0005-2736(83)90284-5.
Using the fluorescent anion 8-anilino-1-naphthalenesulphonate (ANS) for determining the membrane surface potential necessitates that the intrinsic affinity constant Ki for the ANS sites be known. Two methods are presented which do not rely on a determination of Ki at high ionic strength. They are respectively applied to neutral membranes (egg phosphatidylcholine liposomes) and highly charged natural ones (horse bean microsomes and liposomes from their phospholipids). The value of Ki appears to be insensitive to the level of occupancy of the sites, the KCl concentration and the pH in large ranges. Furthermore, the classical Gouy-Chapman model seems to describe correctly the whole set of data, provided apparent mean molecular areas larger than the published crystallographic ones are admitted.
使用荧光阴离子8-苯胺基-1-萘磺酸盐(ANS)测定膜表面电位需要知道ANS位点的固有亲和常数Ki。本文提出了两种不依赖于在高离子强度下测定Ki的方法。它们分别应用于中性膜(卵磷脂脂质体)和高电荷天然膜(蚕豆微粒体及其磷脂脂质体)。在很大范围内,Ki值似乎对位点占据水平、KCl浓度和pH不敏感。此外,经典的 Gouy-Chapman模型似乎能正确描述整个数据集,前提是允许表观平均分子面积大于已发表的晶体学数据。