一种用于测定透化肌纤维中磷酸盐释放的时间分辨测量新方法。
A new method for the time-resolved measurement of phosphate release in permeabilized muscle fibers.
作者信息
Ferenczi M A, He Z H, Chillingworth R K, Brune M, Corrie J E, Trentham D R, Webb M R
机构信息
National Institute for Medical Research, Mill Hill, London, United Kingdom.
出版信息
Biophys J. 1995 Apr;68(4 Suppl):191S-192S; discussion 192S-193S.
Abstract
A new method for the measurement of phosphate release in contracting and relaxed permeabilized muscle fibers is described. The assay is based on a genetically engineered phosphate-binding protein labeled with a coumarin fluorescent probe, which binds inorganic phosphate tightly and shows a fourfold increase in fluorescence upon binding. Measurements of Pi release on the millisecond time scale with sensitivity in the 10 microM range are obtained that provide new information about the relationship between ATP hydrolysis and force production.
摘要
本文描述了一种用于测量收缩和舒张的通透化肌纤维中磷酸盐释放的新方法。该测定基于一种用香豆素荧光探针标记的基因工程化磷酸盐结合蛋白，它能紧密结合无机磷酸盐，并在结合后荧光增强四倍。在毫秒时间尺度上对磷酸盐释放进行测量，灵敏度可达10微摩尔范围，这为ATP水解与力产生之间的关系提供了新信息。
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本文引用的文献
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Biochemistry. 1994 Jul 12;33(27):8262-71. doi: 10.1021/bi00193a013.
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