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采用新型化学发光法以天然底物测定乙酰胆碱酯酶活性。

Determination of acetylcholinesterase activity by a new chemiluminescence assay with the natural substrate.

作者信息

Birman S

出版信息

Biochem J. 1985 Feb 1;225(3):825-8. doi: 10.1042/bj2250825.

DOI:10.1042/bj2250825
PMID:3977858
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1144660/
Abstract

A chemiluminescence method for determining acetylcholinesterase activity is described. It is an adaptation of the chemiluminescence assay of acetylcholine described by Israël & Lesbats [(1981) Neurochem. Int. 3, 81-90; (1981) J. Neurochem. 37, 1475-1483]. The acetylcholinesterase activity is measured by monitoring the increase in light emission produced by the accumulation of choline or by determining the amount of choline generated after a short interval. The assay is rapid and sensitive, and uses the natural substrate of the enzyme. Kinetic data obtained with this procedure for acetylcholinesterase from Torpedo and Electrophorus electric organs were comparable with those obtained by using the method of Ellman, Courtney, Andres & Featherstone [(1961) Biochem. Pharmacol. 7, 88-95]. In addition, it was shown that sodium deoxycholate totally inactivated Torpedo acetylcholinesterase but not the Electrophorus enzyme. Competitive inhibitors of acetylcholinesterase protected the enzyme from inactivation.

摘要

本文描述了一种用于测定乙酰胆碱酯酶活性的化学发光方法。它是对Israël和Lesbats所描述的乙酰胆碱化学发光测定法的改进[(1981)《神经化学国际》3, 81 - 90；(1981)《神经化学杂志》37, 1475 - 1483]。通过监测胆碱积累产生的光发射增加或通过确定短时间间隔后产生的胆碱量来测量乙酰胆碱酯酶活性。该测定法快速且灵敏，并使用该酶的天然底物。用此方法从电鳐和电鳗电器官获得的乙酰胆碱酯酶动力学数据与使用Ellman、Courtney、Andres和Featherstone方法[(1961)《生物化学与药理学》7, 88 - 95]获得的数据相当。此外，已表明脱氧胆酸钠可使电鳐乙酰胆碱酯酶完全失活，但对电鳗酶无此作用。乙酰胆碱酯酶的竞争性抑制剂可保护该酶不被失活。

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本文引用的文献

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The molecular forms of cholinesterase and acetylcholinesterase in vertebrates.脊椎动物体内胆碱酯酶和乙酰胆碱酯酶的分子形式。
Annu Rev Neurosci. 1982;5:57-106. doi: 10.1146/annurev.ne.05.030182.000421.
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Solubilization of membranes by detergents.用去污剂溶解细胞膜。
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