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通过快速蛋白质液相色谱法分离不同形式的S-腺苷甲硫氨酸合成酶

Separation of different forms of S-adenosylmethionine synthetase by fast protein liquid chromatography.

作者信息

van Faassen H, Berger R

机构信息

Department of Pediatrics, University of Groningen, The Netherlands.

出版信息

J Biochem Biophys Methods. 1990 Mar;20(3):189-94. doi: 10.1016/0165-022x(90)90077-p.

Abstract

A method is described by which different forms of S-adenosylmethionine synthetase are separated. The method makes use of fast protein liquid chromatography on an anion exchange hydrophilic polyether resin. Different forms of S-adenosylmethionine synthetase from rat and human liver and kidney and rat zajdela hepatoma cells can be separated within 15 min. From a mixture of rat liver and kidney cytosols all three forms alpha, beta and gamma can be separated. The time needed to separate the different forms of S-adenosylmethionine synthetase is reduced from 3 h with conventional gel filtration methods, to 15 min using this HPLC anion-exchange method. Also the amount of tissue needed to detect the different forms is reduced from 125 mg to 12.5 mg of fresh rat liver tissue. These advantages make this newly developed method applicable when large numbers of samples have to be analyzed or when only small amounts of tissue are available.

摘要

本文描述了一种分离不同形式S-腺苷甲硫氨酸合成酶的方法。该方法利用阴离子交换亲水性聚醚树脂进行快速蛋白质液相色谱。来自大鼠和人类肝脏、肾脏以及大鼠Zajdela肝癌细胞的不同形式的S-腺苷甲硫氨酸合成酶可在15分钟内分离。从大鼠肝脏和肾脏细胞溶质的混合物中,α、β和γ这三种形式都可以被分离出来。分离不同形式的S-腺苷甲硫氨酸合成酶所需的时间从传统凝胶过滤方法的3小时减少到使用这种高效液相色谱阴离子交换方法的15分钟。而且检测不同形式所需的组织量从125毫克新鲜大鼠肝脏组织减少到12.5毫克。这些优点使得这种新开发的方法适用于需要分析大量样品或仅有少量组织可用的情况。

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