一种快速灵敏的测定核糖体失活蛋白酶活性的方法。
A rapid and sensitive method to measure the enzymatic activity of ribosome-inactivating proteins.
作者信息
Brigotti M, Barbieri L, Valbonesi P, Stirpe F, Montanaro L, Sperti S
机构信息
Dipartimento di Patologia sperimentale dell'Università degli Studi di Bologna, Via San Giacomo 14, I-40126 Bologna, Italy.
出版信息
Nucleic Acids Res. 1998 Sep 15;26(18):4306-7. doi: 10.1093/nar/26.18.4306.
Abstract
A method is described in which the adenosine- N -glycosidase activity of ribosome-inactivating proteins (RIPs) is measured using as substrate a 2251 bp [3H]DNA obtained by PCR amplification of the 731-2981 region of the pBR322 plasmid. The DNA, labelled in the purine ring of adenine, proved a good substrate for all three RIPs tested (PAP-S, ricin and shiga-like toxin I). The method, which measures directly the [3H]adenine released, is highly specific, extremely rapid and quantitative in a wide range of RIP concentrations.
摘要
本文描述了一种方法，其中核糖体失活蛋白（RIPs）的腺苷-N-糖苷酶活性通过使用经PCR扩增pBR322质粒731-2981区域获得的2251 bp [³H]DNA作为底物来测量。该DNA在腺嘌呤的嘌呤环中标记，被证明是所有三种测试的RIPs（PAP-S、蓖麻毒素和志贺样毒素I）的良好底物。该方法直接测量释放的[³H]腺嘌呤，在广泛的RIP浓度范围内具有高度特异性、极快且定量。
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本文引用的文献
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The RNA-N-glycosidase activity of Shiga-like toxin I: kinetic parameters of the native and activated toxin.志贺样毒素I的RNA-N-糖苷酶活性：天然毒素和活化毒素的动力学参数
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