用于测量膜蛋白旋转扩散的荧光三重态探针。
Fluorescent triplet probes for measuring the rotational diffusion of membrane proteins.
作者信息
Johnson P, Garland P B
出版信息
Biochem J. 1982 Apr 1;203(1):313-21. doi: 10.1042/bj2030313.
Abstract
We have previously described a method for measuring the rotational diffusion of membrane proteins by using fluorescent triplet probes [Johnson & Garland (1981) FEBS Lett. 135, 252-256]. We now describe the criteria by which the suitability of such probes may be judged. In general, the greatest sensitivity is achievable with probes where the ratio of the quantum yields for prompt fluorescene (phi f) and triplet formation (phi t) are high, as with Rhodamine (phi f/phi t congruent to 10(3)). However, considerations of heat generation at the sample membrane, of time resolution of fast rotations and of irreversible bleaching of the fluorescent probe also apply. The immediate environment of a probe molecule at a membrane protein must also be important in determining the performance of a given probe. Nevertheless, we describe guidelines for evaluating the likely usefulness of fluorescent triplet probes in measurements of membrane protein rotation.
摘要
我们之前描述了一种通过使用荧光三重态探针来测量膜蛋白旋转扩散的方法[约翰逊和加兰（1981年），《欧洲生物化学学会联合会快报》135卷，252 - 256页]。我们现在描述判断此类探针适用性的标准。一般来说，对于那些即时荧光（φf）和三重态形成（φt）的量子产率之比很高的探针，能实现最大灵敏度，比如罗丹明（φf/φt约为10³）。然而，样品膜处的发热、快速旋转的时间分辨率以及荧光探针的不可逆漂白等因素也需要考虑。探针分子在膜蛋白处的直接环境对于确定给定探针的性能也必定很重要。尽管如此，我们还是描述了评估荧光三重态探针在膜蛋白旋转测量中可能有用性的指导原则。
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