使用用N-(1-芘基)碘乙酰胺标记的肌动蛋白来研究肌动蛋白与肌球蛋白亚片段以及肌钙蛋白/原肌球蛋白之间的相互作用。
The use of actin labelled with N-(1-pyrenyl)iodoacetamide to study the interaction of actin with myosin subfragments and troponin/tropomyosin.
作者信息
Criddle A H, Geeves M A, Jeffries T
出版信息
Biochem J. 1985 Dec 1;232(2):343-9. doi: 10.1042/bj2320343.
Abstract
A pyrene label attached to Cys-374 of actin has been shown to be a useful probe for monitoring the interaction of actin with myosin subfragments [Kouyama & Mihashi (1981) Eur. J. Biochem. 114, 33-38]. We report that the presence of this label decreases the affinity of actin for myosin subfragment 1 by less than a factor of 2. The rate of actin binding is unaffected by the label and the dissociation rate is increased by up to a factor of 2. Both the rate of actin binding to, and the rate of actin dissociation from, heavy meromyosin show two phases when monitored by pyrene fluorescence. Thin filiments reconstituted from pyrene-labelled actin show a 5% increase in pyrene fluorescence on binding Ca2+.
摘要
已证明附着于肌动蛋白Cys-374的芘标签是监测肌动蛋白与肌球蛋白亚片段相互作用的有用探针[小山和三桥（1981年），《欧洲生物化学杂志》114卷，33 - 38页]。我们报告称，该标签的存在使肌动蛋白对肌球蛋白亚片段1的亲和力降低不到2倍。肌动蛋白结合速率不受该标签影响，而解离速率最多增加2倍。当通过芘荧光监测时，肌动蛋白与重酶解肌球蛋白的结合速率和解离速率均呈现两个阶段。由芘标记的肌动蛋白重构的细肌丝在结合Ca2+时芘荧光增加5%。
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The use of actin labelled with N-(1-pyrenyl)iodoacetamide to study the interaction of actin with myosin subfragments and troponin/tropomyosin.使用用N-(1-芘基)碘乙酰胺标记的肌动蛋白来研究肌动蛋白与肌球蛋白亚片段以及肌钙蛋白/原肌球蛋白之间的相互作用。
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The effect of EDTA on spectral properties of ATP-, ADP-, and ITP-G-actin.乙二胺四乙酸（EDTA）对ATP-、ADP-和ITP-G-肌动蛋白光谱特性的影响。
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