PHALLOTOXIN AND ACTIN BINDING ASSAY BY FLUORESCENCE ENHANCEMENT
PHALLOTOXIN AND ACTIN BINDING ASSAY BY FLUORESCENCE ENHANCEMENT
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DOI：
10.1016/0003-2697(92)90299-m
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发表时间：
1992-01-01
影响因子：
2.9
通讯作者：
HAUGLAND, RP
中科院分区：
文献类型：
--
作者：
HUANG, ZJ;HAUGLAND, RP;HAUGLAND, RP
在聚合缓冲液中与F-肌动蛋白结合时，发现与鬼臼毒素结合的五个荧光团的荧光被特异性地增强。罗丹明鬼臼糖苷的荧光增强作用最强，为9倍。肌动蛋白对罗丹明菲洛定的荧光滴定符合化学计量结合规律。罗丹明鬼臼苷在5μm处的荧光增强与F-肌动蛋白浓度在2μm以下呈线性关系，因此可作为F-肌动蛋白定量的一种简便方法。在竞争分析中，其他鬼臼毒素降低了罗丹明鬼臼毒素与聚合肌动蛋白结合所产生的荧光增强。这种还原还允许方便地测量任何竞争的鬼臼毒素的结合常数。
The fluorescence of five fluorophores conjugated to phallotoxins was found to be specifically enhanced upon binding to F-actin in a polymerizing buffer. Rhodamine phalloidin had the greatest fluorescence enhancement of ninefold. The fluorescence titration of rhodamine phalloidin by actin was shown to be consistent with stoichiometric binding. The fluorescence enhancement of rahodamine phalloidin at 5 μm is linearly related to F-actin concentrations up to 2 μm and therefore can be used as an easy means of F-actin quantitation. In a competition assay, other phallotoxins reduce the fluorescence enhancement that results from the binding of rhodamine phalloidin to polymerized actin. This reduction also permits a convenient measurement of the binding constants of any competing phallotoxins.