PHALLOTOXIN AND ACTIN BINDING ASSAY BY FLUORESCENCE ENHANCEMENT

被引：80
作者：
HUANG, ZJ
HAUGLAND, RP
YOU, WM
HAUGLAND, RP
机构：
[1] Molecular Probes, Inc., Eugene, OR 97402
来源：
ANALYTICAL BIOCHEMISTRY | 1992年 / 200卷 / 01期
关键词：
D O I：
10.1016/0003-2697(92)90299-M
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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. © 1992.
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页码：199 / 204
页数：6
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